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Methionine

Here is the technical story about
The Amino Acid
Methionine

Methionine is an amino acid and probably is the second most important ingredient in Life Glow.  There is 300 mg of Methionine in Life Glow, and also in Super Life Glow.

me·thi·o·nine (me-thì¹e-nên´) noun

A sulfur-containing essential amino acid, obtained from various proteins or prepared synthetically and used as a dietary supplement and in pharmaceuticals.

This amino acid has the ability to help keep the inside walls of the arteries naturally clear of accumulated fatty deposits.

The actual "stuff" which it helps create is an enzyme called "lipoprotein lipase."  This enzyme is what actually does the artery cleaning.

lipoprotein

Definition:  a combination of a lipid (fat) and protein, possessing the general properties (e.g., solubility) of proteins.  Practically all of the lipids of the plasma are present as lipoprotein complexes, a- and B-lipoproteins being distinguished by electrophoresis.  The B-lipoproteins transport more of the total plasma cholesterol, contain a higher concentration of both free and esterfied cholesterol, and have a higher cholesterol/phospholipid ratio than a-lipoproteins.

lipoprotein lipase

lipase:  any of a group of widely occurring enzymes that catalyze the hydrolysis of ester linkages between the fatty acids and glycerol of the triglycerides and phospholipids. 

lipoprotein lipase:  an esterase (enzyme) that catalyzes (speeds up or causes) the hydrolysis (using water to breakdown) of the constituent triglycerides (a common type of fat in humans) of chylomicrons (a mixture of fats, with a small amount of protein, in the form that occurs in the intestine and also, then, in the blood) to form free fatty acid anions (negatively charged particles) and glycerol (one of the components of the breakdown of a fat).

apoprotein:  This is a substance which activates the lipoprotein lipase.

Here is a much less formal explanation of "lipoprotein lipase:"

Lipoprotein Lipase (LPL):  This is an enzyme which helps remove fats (cholesterol) which might be deposited on the inside walls of the arteries.

It turns out that the amino acid, methionine, causes an increase of LPL in the body, and thus methionine can help the body get rid of excess cholesterol on the inside of the arteries.

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HealthGate Documents

Record 1 from database: MEDLINE
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Title

Recombinant human tumour necrosis factor-alpha suppresses synthesis, activity and secretion of lipoprotein lipase in cultures of a human osteosarcoma cell line.

Author

Sakayama K; Masuno H; Okumura H; Shibata T; Okuda H

Address

Department of Orthopaedic Surgery, School of Medicine, Ehime University, Japan.

Source

Biochem J, 1996 Jun, 316 ( Pt 3):, 813-7

Abstract

The effect of recombinant human tumour necrosis factor-alpha (TNF-alpha) on synthesis, activity and secretion of lipoprotein lipase (LPL) was examined using a human osteosarcoma cell line, osteosarcoma Takase (OST). Treatment of OST cells with TNF-alpha decreased LPL synthesis, resulting in a decrease in expression of activity and secretion of LPL. When OST cells were incubated with glycerol tri[1-14C]palmitate, TNF-alpha decreased dose- and time-dependently the production of 14CO2 and the amounts of radioactivity incorporated into cellular triacylglycerol and phospholipid. The similar reduction of synthesis and activity of LPL as suppression of CO2 production and cellular lipid synthesis indicated that the suppression of 14CO2 production and 14C-labelled lipid synthesis was secondary. TNF-alpha also suppressed expression of proliferating cell nuclear antigen, indicating that it had an anti-proliferative activity on OST cells. The findings suggest that one cause of the anti-proliferative activity of TNF-alpha is the suppression of the LPL-mediated supply of non-esterified fatty acids as an energy source for growth.

Language of Publication

English

Unique Identifier

96265046

MeSH Heading (Major)

Lipids|*BI; Lipoprotein Lipase|BI/*ME/SE; Tumor Necrosis Factor|*PD

MeSH Heading

Bone Neoplasms; Carbon Dioxide|AN; Carbon Radioisotopes; Cell Line; Diglycerides|BI; Fatty Acids, Nonesterified|BI; Human; Kinetics; Methionine|ME; Osteosarcoma; Phospholipids|BI; Recombinant Proteins|PD; Sulfur Radioisotopes; Triglycerides|BI/ME; Tumor Cells, Cultured

Publication Type

JOURNAL ARTICLE

ISSN

0264-6021

Country of Publication

ENGLAND

Record 2 from database: MEDLINE
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Title

Isolation and characterization of recombinant human apolipoprotein C-II expressed in Escherichia coli.

Author

Wang CS; Downs D; Dashti A; Jackson KW

Address

Protein Studies Program, Oklahoma Medical Research Foundation, Oklahoma City 73104, USA. chi-sun-wang@omrf.uokhsc.edu

Source

Biochim Biophys Acta, 1996 Aug, 1302:3, 224-30

Abstract

A full-length recombinant human apolipoprotein C-II (ApoC-II) has been successfully expressed in Escherichia coli using the T7 expression system. The recombinant ApoC-II. which was expressed intracellularly in the inclusion bodies, was solubilized with 8 M urea and purified using Sephadex G-75 gel permeation chromatography. Four liters of the bacterial culture yielded 16-20 mg of purified recombinant ApoC-II. Sequencing and mass spectrometric analyses indicated that the isolated recombinant ApoC-II contained predominantly (64%) the native form with threonine as the N-terminus, but also contained a minor (36%) molecular form of ApoC-II with an additional methionine at the N-terminus (Met-ApoC-II). Analysis of the recombinant ApoC-II by tryptic digestion and high performance liquid chromatography-electrospray mass spectrometry provides additional conclusive evidence that, with the exception of the N-terminus of Met-ApoC-II, the expressed ApoC-II has the expected peptide sequence. However, this extra N-terminal methionine residue can be excised by further in vitro treatment with methionine aminopeptidase. The purified recombinant ApoC-II was found to be competent in the activation of bovine milk lipoprotein lipase. Thus, the recombinant ApoC-II prepared from E. coli may have a pharmacological application for the treatment of patients with genetic hypertriglyceridemia caused by ApoC-II deficiency.

Language of Publication

English

Unique Identifier

96350451

MeSH Heading (Major)

Apolipoproteins C|CH/GE/*IP; Escherichia coli|*GE

MeSH Heading

Amino Acid Sequence; Aminopeptidases|ME; Animal; Base Sequence; Cattle; Chromatography, Gel; Chromatography, High Pressure Liquid; Electrophoresis, Polyacrylamide Gel; Enzyme Activation|DE; Human; Lipoprotein Lipase|ME; Milk|EN; Molecular Sequence Data; Recombinant Proteins|CH/IP/PD; Spectrum Analysis, Mass; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

0006-3002

Country of Publication

NETHERLANDS

Record 3 from database: MEDLINE
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Title

Molecular characterization of human hepatic lipase deficiency. In vitro expression of two naturally occurring mutations.

Author

Durstenfeld A; Ben Zeev O; Reue K; Stahnke G; Doolittle MH

Address

Lipid Research, Veterans Administration Wadsworth Medical Center, Los Angeles, CA 90073.

Source

Arterioscler Thromb, 1994 Mar, 14:3, 381-5

Abstract

Individuals with hepatic lipase (HL) deficiency are often characterized by elevated levels of triglycerides and cholesterol and may be subject to premature atherosclerosis. Missense mutations in the HL gene have been identified in two affected families: substitutions of serine for phenylalanine at amino acid 267 and threonine for methionine at amino acid 383 (S267F and T383M, respectively). To confirm the role of S267F and T383M, respectively). To confirm the role of mutations separately into human HL cDNA by site-directed mutagenesis, and the resulting constructs were independently expressed in COS cells. HL activity and mass were measured and compared with wild-type HL transfectants to determine the effect of these mutations on lipase activity and secretion. Although similar amounts of HL protein were detected intracellularly after transfection with the wild-type and mutant constructs, S267F and T383M HL activity levels were markedly decreased: in S267F, no HL activity was detected, and activity levels in T383M were 38% of wild-type HL. Heparin-induced secretion of the two HL mutants was also severely affected: no detectable activity could be measured in the media of S267F, although some inactive mass (12% of wild-type HL) was secreted; mutant T383M secreted 4% and 20% of wild-type activity and mass, respectively. These results indicate that the single amino acid substitution present in HL S267F is sufficient to render the enzyme completely nonfunctional; in contrast, the T383M mutant retains partial activity but is poorly secreted. Thus, these defects appear capable of accounting for the HL-deficient phenotypes exhibited by individuals carrying the T383M and S267F mutations.

Language of Publication

English

Unique Identifier

94169141

MeSH Heading (Major)

Lipase|*DF/GE; Liver|*EN

MeSH Heading

Base Sequence; Human; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Support, Non-U.S. Gov't; Support, U.S. Gov't, Non-P.H.S.; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

1049-8834

Country of Publication

UNITED STATES

Record 4 from database: MEDLINE
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Title

Apo B100-containing lipoproteins are secreted by the heart.

Author

Borén J; Véniant MM; Young SG

Address

Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, California 94141-9100, USA. jboren@gladstone.ucsf.edu

Source

J Clin Invest, 1998 Mar, 101:6, 1197-202

Abstract

The apo B gene is expressed in the human heart and in the hearts of human apo B transgenic mice generated with large genomic clones spanning the human apo B gene. [35S]Methionine metabolic labeling experiments demonstrated that apo B100-containing lipoproteins are secreted by human heart tissue and by human apo B transgenic and nontransgenic mouse heart tissue. Density gradient analysis revealed that most of the secreted heart lipoproteins were LDLs, even when the labeling experiments were performed in the presence of tetrahydrolipstatin, an inhibitor of lipoprotein lipase. Western blots with a microsomal triglyceride transfer protein) (MTP)-specific antiserum demonstrated that the microsomes of the heart contain the 97-kD subunit of MTP (the subunit involved in the transfer of lipids and assembly of lipoproteins). Metabolic labeling of mouse heart tissue in the presence of BMS-192951, an MTP inhibitor, abolished lipoprotein secretion by the heart but resulted in the secretion of two apo B proteolytic fragments (80 and 120 kD), which were found in the bottom fraction of the density gradient. These studies reveal that the heart, and not just the liver and intestine, secretes apo B-containing lipoproteins. We speculate that lipoprotein secretion by the heart represents a mechanism for removing excess lipids from the heart.

Language of Publication

English

Unique Identifier

98171505

MeSH Heading (Major)

Apolipoproteins B|*GE/ME/*SE; Myocardium|*ME

MeSH Heading

Animal; Blotting, Western; Carrier Proteins|AI/IM/ME; Centrifugation, Density Gradient; Enzyme Inhibitors|PD; Fatty Acids|ME; Gene Expression; Human; Intestines|ME; Lactones|PD; Lipoprotein Lipase|AI; Lipoproteins|AN/ME/SE; Lipoproteins, LDL|AN/ME/SE; Liver|SE; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microsomes|ME; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

0021-9738

Country of Publication

UNITED STATES

Record 5 from database: MEDLINE
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Title

Endothelial cells synthesize and process apolipoprotein B.

Author

Sivaram P; Vanni Reyes T; Goldberg IJ

Address

Division of Preventive Medicine and Nutrition, Department of Medicine, Columbia University, New York, New York 10032, USA.

Source

J Biol Chem, 1996 Jun, 271:25, 15261-6

Abstract

We reported previously that a 116-kDa lipoprotein lipase (LPL)-binding protein from endothelial cells has sequence homology to the amino-terminal region of apolipoprotein (apo) B. We now tested whether endothelial cells synthesize apoB mRNA and protein. Primers were designed to the human apoB cDNA sequence and reverse transcription polymerase chain reaction was performed using total RNA isolated from bovine and human endothelial cells. With primers to the 5' region of the apoB mRNA (amino-terminal region of apoB protein) expected size PCR products were generated from both bovine and human endothelial cells as well as from mouse liver RNA, which was used as a control. Primers designed to the 3' region of apoB mRNA generated PCR products from human endothelial cells and HepG2 cells but not from bovine or mouse cells. These data suggest that endothelial cells contain full-length apoB mRNA and that the 5' or the amino-terminal region of apoB is highly conserved from mouse to human. This was confirmed by direct sequencing of the mouse and bovine PCR products. To test whether apoB protein was produced, bovine endothelial cell proteins were metabolically labeled with [35S]methionine/cysteine or [3H]leucine and immunoprecipitated with anti-human apoB antibodies. Using extracts from cells labeled for 1 h, monoclonal antibody 47, directed to the low density lipoprotein receptor binding region of apoB, precipitated a protein of approximate molecular mass 550,000, the size of full-length apoB. Immunoprecipitation of the 550-kDa protein was abolished in the presence of added unlabeled low density lipoprotein. From cells labeled for 16 h, a 116-kDa protein was immunoprecipitated by polyclonal anti-apoB antibodies. This protein was partly released from cells by heparin treatment. Pulse-chase analysis showed that the 116-kDa fragment appeared at the same time as the full-length apoB began disappearing. The immunoprecipitated 116-kDa fragment also bound labeled LPL on ligand blot, further suggesting that it is an amino-terminal fragment of apoB. Incubation of endothelial cells with oleic acid (0.25 and 0.5 mM) did not significantly alter the production of either the full-length apoB or the 116-kDa fragment. These data show that endothelial cells synthesize apoB. The full-length apoB appears to be cleaved to form a 116-kDa fragment that can function as a LPL-binding protein.

Language of Publication

English

Unique Identifier

96279028

MeSH Heading (Major)

Apolipoproteins B|*BI/CH; Endothelium, Vascular|DE/*ME

MeSH Heading

Amino Acid Sequence; Animal; Aorta; Base Sequence; Carcinoma, Hepatocellular; Cattle; Cell Line; Cells, Cultured; Conserved Sequence; DNA Primers; DNA, Complementary; Gene Library; Human; Intestines; Kinetics; Lipoprotein Lipase|ME; Liver Neoplasms; Mice; Models, Biological; Molecular Sequence Data; Oleic Acids|PD; Support, U.S. Gov't, P.H.S.; Tumor Cells, Cultured

Publication Type

JOURNAL ARTICLE

ISSN

0021-9258

Country of Publication

UNITED STATES

Record 6 from database: MEDLINE
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Title

Regulation of lipoprotein lipase translation by epinephrine in 3T3-L1 cells. Importance of the 3' untranslated region.

Author

Yukht A; Davis RC; Ong JM; Ranganathan G; Kern PA

Address

Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California 90048, USA.

Source

J Clin Invest, 1995 Nov, 96:5, 2438-44

Abstract

Lipoprotein lipase (LPL) is a central enzyme in lipoprotein metabolism and is in part responsible for adipocyte lipid accumulation. Catecholamines are known to decrease the activity of LPL in adipocytes, and we have previously demonstrated that this inhibition occurs posttranscriptionally, with a prominent inhibition of LPL translation. To better characterize the inhibition of LPL translation, 3T3-L1 cells were differentiated into adipocytes, and exposed to epinephrine. Epinephrine induced a dose-dependent decrease in LPL synthesis using [35S]methionine incorporation, with no change in LPL mRNA levels, demonstrating translational regulation of LPL in this cell line. The poly A-enriched RNA from epinephrine-treated cells was translated well in vitro, and there was no difference in the polysome profiles from control and epinephrine-treated cells, suggesting that epinephrine did not affect mRNA editing, and did not induce an inhibition of translation initiation. To obtain evidence for the presence of an inhibitory factor, a cytoplasmic extract from control, and epinephrine-treated adipocytes was human. When compared to the control cell extract, the epinephrine-treated cell extract sharply inhibited LPL translation in vitro, yet had no effect on the translation of other mRNAs. Epinephrine-treated cells had fourfold more of this inhibitor activity than control cells, and this translation inhibition was partially reversed by heat treatment. To determine what region of the LPL mRNA was involved in the translation inhibition, different LPL constructs were synthesized. The inhibitory effect of the epinephrine-treated cell extract was dependent on the presence of the first 40 nucleotides of the 3' (untranslated region UTR) (nucleotides 1599-1638), whereas deletion of the 5' UTR and other areas of the 3' UTR had no effect on translation inhibition. When a sense RNA strand corresponding to this region was added to the in vitro translation reaction, it restored translation towards normal, suggesting that the sense strand was competing for a transacting binding protein. Thus, epinephrine-treated adipocytes produced a transacting factor, probably a protein, that interacted with a region on the LPL mRNA between nucleotides 1599 and 1638, resulting in an inhibition of translation. These studies add new insight into the hormonal regulation of LPL.

Language of Publication

English

Unique Identifier

96066711

MeSH Heading (Major)

Adipocytes|*EN; Lipoprotein Lipase|*BI; RNA, Messenger|GE/*ME; Translation, Genetic|*DE

MeSH Heading

Adrenergic Agonists|PD; Animal; Base Sequence; Dose-Response Relationship, Drug; Epinephrine|PD; Human; Mice; Molecular Sequence Data; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; 3T3 Cells

Publication Type

JOURNAL ARTICLE

ISSN

0021-9738

 

---

HealthGate Documents

Record 7 from database: MEDLINE
Return To Top

Title

Apo B100-containing lipoproteins are secreted by the heart.

Author

Borén J; Véniant MM; Young SG

Address

Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, California 94141-9100, USA. jboren@gladstone.ucsf.edu

Source

J Clin Invest, 1998 Mar, 101:6, 1197-202

Abstract

The apo B gene is expressed in the human heart and in the hearts of human apo B transgenic mice generated with large genomic clones spanning the human apo B gene. [35S]Methionine metabolic labeling experiments demonstrated that apo B100-containing lipoproteins are secreted by human heart tissue and by human apo B transgenic and nontransgenic mouse heart tissue. Density gradient analysis revealed that most of the secreted heart lipoproteins were LDLs, even when the labeling experiments were performed in the presence of tetrahydrolipstatin, an inhibitor of lipoprotein lipase. Western blots with a microsomal triglyceride transfer protein) (MTP)-specific antiserum demonstrated that the microsomes of the heart contain the 97-kD subunit of MTP (the subunit involved in the transfer of lipids and assembly of lipoproteins). Metabolic labeling of mouse heart tissue in the presence of BMS-192951, an MTP inhibitor, abolished lipoprotein secretion by the heart but resulted in the secretion of two apo B proteolytic fragments (80 and 120 kD), which were found in the bottom fraction of the density gradient. These studies reveal that the heart, and not just the liver and intestine, secretes apo B-containing lipoproteins. We speculate that lipoprotein secretion by the heart represents a mechanism for removing excess lipids from the heart.

Language of Publication

English

Unique Identifier

98171505

MeSH Heading (Major)

Apolipoproteins B|*GE/ME/*SE; Myocardium|*ME

MeSH Heading

Animal; Blotting, Western; Carrier Proteins|AI/IM/ME; Centrifugation, Density Gradient; Enzyme Inhibitors|PD; Fatty Acids|ME; Gene Expression; Human; Intestines|ME; Lactones|PD; Lipoprotein Lipase|AI; Lipoproteins|AN/ME/SE; Lipoproteins, LDL|AN/ME/SE; Liver|SE; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microsomes|ME; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

0021-9738

Country of Publication

UNITED STATES

Record 8 from database: MEDLINE
Return To Top

Title

Endothelial cells synthesize and process apolipoprotein B.

Author

Sivaram P; Vanni Reyes T; Goldberg IJ

Address

Division of Preventive Medicine and Nutrition, Department of Medicine, Columbia University, New York, New York 10032, USA.

Source

J Biol Chem, 1996 Jun, 271:25, 15261-6

Abstract

We reported previously that a 116-kDa lipoprotein lipase (LPL)-binding protein from endothelial cells has sequence homology to the amino-terminal region of apolipoprotein (apo) B. We now tested whether endothelial cells synthesize apoB mRNA and protein. Primers were designed to the human apoB cDNA sequence and reverse transcription polymerase chain reaction was performed using total RNA isolated from bovine and human endothelial cells. With primers to the 5' region of the apoB mRNA (amino-terminal region of apoB protein) expected size PCR products were generated from both bovine and human endothelial cells as well as from mouse liver RNA, which was used as a control. Primers designed to the 3' region of apoB mRNA generated PCR products from human endothelial cells and HepG2 cells but not from bovine or mouse cells. These data suggest that endothelial cells contain full-length apoB mRNA and that the 5' or the amino-terminal region of apoB is highly conserved from mouse to human. This was confirmed by direct sequencing of the mouse and bovine PCR products. To test whether apoB protein was produced, bovine endothelial cell proteins were metabolically labeled with [35S]methionine/cysteine or [3H]leucine and immunoprecipitated with anti-human apoB antibodies. Using extracts from cells labeled for 1 h, monoclonal antibody 47, directed to the low density lipoprotein receptor binding region of apoB, precipitated a protein of approximate molecular mass 550,000, the size of full-length apoB. Immunoprecipitation of the 550-kDa protein was abolished in the presence of added unlabeled low density lipoprotein. From cells labeled for 16 h, a 116-kDa protein was immunoprecipitated by polyclonal anti-apoB antibodies. This protein was partly released from cells by heparin treatment. Pulse-chase analysis showed that the 116-kDa fragment appeared at the same time as the full-length apoB began disappearing. The immunoprecipitated 116-kDa fragment also bound labeled LPL on ligand blot, further suggesting that it is an amino-terminal fragment of apoB. Incubation of endothelial cells with oleic acid (0.25 and 0.5 mM) did not significantly alter the production of either the full-length apoB or the 116-kDa fragment. These data show that endothelial cells synthesize apoB. The full-length apoB appears to be cleaved to form a 116-kDa fragment that can function as a LPL-binding protein.

Language of Publication

English

Unique Identifier

96279028

MeSH Heading (Major)

Apolipoproteins B|*BI/CH; Endothelium, Vascular|DE/*ME

MeSH Heading

Amino Acid Sequence; Animal; Aorta; Base Sequence; Carcinoma, Hepatocellular; Cattle; Cell Line; Cells, Cultured; Conserved Sequence; DNA Primers; DNA, Complementary; Gene Library; Human; Intestines; Kinetics; Lipoprotein Lipase|ME; Liver Neoplasms; Mice; Models, Biological; Molecular Sequence Data; Oleic Acids|PD; Support, U.S. Gov't, P.H.S.; Tumor Cells, Cultured

Publication Type

JOURNAL ARTICLE

ISSN

0021-9258

Country of Publication

UNITED STATES

Record 9 from database: MEDLINE
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Title

Identification of a neutral lipid core in a transiently expressed and secreted lipoprotein containing an apoB-48-like apolipoprotein.

Author

Spring DJ; Lee SM; Puppione DL; Phillips M; Elovson J; Schumaker VN

Address

Department of Chemistry and Biochemistry, University of California, Los Angeles.

Source

J Lipid Res, 1992 Feb, 33:2, 233-40

Abstract

The presence of core lipids in lipoproteins expressed and secreted by transfected HepG2 cells was demonstrated by measuring the densities of these lipoproteins before and after treatment with a bacterial lipase specific for neutral lipids. HepG2 cells were reproducibly transfected with pRSV/B48, containing a truncated human apolipoprotein B-100 (apoB-100) cDNA (nucleotides 1 to 6860, where nucleotide 129 is the start of translation). Northern blots of cellular message probed with apoB-48 showed abundant transcription of an apoB-48-sized message as well as endogenous apoB-100 message. When grown in the presence of [35S]methionine, pRSV/B48-transfected cells secreted lipoproteins containing an apoB-48-like apolipoprotein. This lipoprotein banded at a density of 1.11 g/ml in isopycnic NaBr gradients. Electron microscopy of the apoB-48-containing lipoproteins demonstrated spherical particles with an average diameter of 124A. A sedimentation rate of 8.4S was measured by sucrose gradient sedimentation. When the apoB-48-containing particles were treated with a bacterial lipase (from Chromobacterium viscosum), shown to hydrolyze triglycerides and cholesteryl esters but not phospholipids, their density increased to 1.18 g/ml, consistent with removal of core lipids. When the secreted lipoprotein was modeled as a spherical particle containing a single molecule of apoB-48, a triglyceride-filled core, and a surface monolayer of phospholipid and protein, the hydrodynamic properties were consistent with the observed sedimentation coefficient, buoyant densities before and after lipase treatment, and the diameter as seen with the electron microscope. These data indicate that transfected HepG2 cells assembled and secreted lipoproteins possessing the same physical structure as naturally occurring lipoproteins.

Language of Publication

English

Unique Identifier

92235556

MeSH Heading (Major)

Apolipoproteins B|*CH/GE; Lipids|*CH; Lipoproteins|*CH/GE/SE/UL

MeSH Heading

Blotting, Northern; Chromobacterium|EN; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Human; Lipase|ME; Microscopy, Electron; Plasmids; Precipitin Tests; Support, U.S. Gov't, Non-P.H.S.; Support, U.S. Gov't, P.H.S.; Tumor Cells, Cultured; Ultracentrifugation

Publication Type

JOURNAL ARTICLE

ISSN

0022-2275

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.3 (Lipase); 0 (apolipoprotein B-48); 0 (Apolipoproteins B); 0 (Lipids); 0 (Lipoproteins); 0 (Plasmids)

Record 10 from database: MEDLINE
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Title

Regulation of hepatic triglyceride lipase by thyroid hormone in HepG2 cells.

Author

Kihara S; Wölle J; Ehnholm C; Chan L; Oka K

Address

Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030.

Source

J Lipid Res, 1993 Jun, 34:6, 961-70

Abstract

Hypothyroidism has been reported to be associated with reduced hepatic triglyceride lipase (HTGL) activity. In order to understand the molecular mechanism by which thyroid hormone regulates HTGL activity, effects of triiodothyronine (T3) on HTGL activity, mRNA level, transcription run-on activity, and protein synthetic rate were studied in HepG2 cells. HepG2 cells treated with 1 nM T3 showed an increase in HTGL activity that was first detected at 24 h; HTGL activity continued to increase at 36 h and stayed at the elevated level at 48 and 60 h. At maximal stimulation (48 h), T3-treated cells had the following HTGL activities: 155% in spontaneously released (SR) and 224% in heparin-releasable (HR) HTGL activities (mean levels compared to control). Stimulation of HTGL activity by T3 was dose-dependent and saturable. There was, however, no change in HTGL mRNA level throughout the course of T3 treatment. The effects of T3 were reduced when transcription was blocked by actinomycin D (mean level compared to actinomycin D treatment in the absence of T3: 109% in SR and 127% in HR activities) or translation was blocked by cycloheximide (127% in SR and 122% in HR activities), but HTGL activities were still significantly higher than control. Nuclear run-on assays indicate that T3 did not change the rate of transcription of the HTGL gene. We further determined the rate of HTGL synthesis by measuring the amount of [35S]methionine incorporated into newly synthesized HTGL immunoprecipitated by a monospecific anti-human HTGL antibody. We found that the T3-stimulated increase in HTGL activity was not accompanied by any change in the rate of HTGL biosynthesis. Our experimental data indicate that the T3 stimulation of HTGL activity in HepG2 cells is mediated at posttranscriptional and posttranslational levels. The partial but significant inhibition of the T3 stimulation of HTGL activity by actinomycin D and cycloheximide suggests that the effects of T3 may be mediated by other cellular processes that are more directly regulated by the hormone. This study represents the initial report on the mechanism of HTGL activation by physiological concentrations of thyroid hormone.

Language of Publication

English

Unique Identifier

93359820

MeSH Heading (Major)

Lipase|*ME; Liver|*EN; Triiodothyronine|*PH

MeSH Heading

Base Sequence; Gene Expression Regulation, Enzymologic; Human; Molecular Sequence Data; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; Tumor Cells, Cultured

Publication Type

JOURNAL ARTICLE

ISSN

0022-2275

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.3 (Lipase); 6893-02-3 (Triiodothyronine)

Record 11from database: MEDLINE
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Title

A hepatic lipase gene mutation associated with heritable lipolytic deficiency.

Author

Hegele RA; Vezina C; Moorjani S; Lupien PJ; Gagne C; Brun LD; Little JA; Connelly PW

Address

Department of Medicine, St. Michael's Hospital, University of Toronto, Ontario, Canada.

Source

J Clin Endocrinol Metab, 1991 Mar, 72:3, 730-2

Abstract

Absent hepatic lipase (HL) activity results in dyslipidemia and premature atherosclerosis. DNA sequencing of the HL gene from subjects with heritable HL deficiency identified a new C to T substitution within exon 8 that in the mature enzyme caused a threonine to methionine change at position 383 (T383M). With a rapid DNA detection method we observed that all 6 individuals with complete HL deficiency from 2 families had the T383M mutation. None of 50 random unrelated unaffected subjects had this mutation. We propose that T383M is specific to families with heritable HL deficiency. Furthermore, structural variation at the HL gene, possibly in combination with other factors, appears to be etiologic in HL deficiency.

Language of Publication

English

Unique Identifier

91147553

MeSH Heading (Major)

Lipase|*GE; Liver|*EN

MeSH Heading

Adult; Base Sequence; Exons; Female; Genotype; Human; Male; Middle Age; Molecular Sequence Data; Mutation; Polymorphism, Restriction Fragment Length

Publication Type

JOURNAL ARTICLE

ISSN

0021-972X

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.3 (Lipase)

Record 12 from database: MEDLINE
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Title

Regulation of lipoprotein lipase translation by epinephrine in 3T3-L1 cells. Importance of the 3' untranslated region.

Author

Yukht A; Davis RC; Ong JM; Ranganathan G; Kern PA

Address

Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California 90048, USA.

Source

J Clin Invest, 1995 Nov, 96:5, 2438-44

Abstract

Lipoprotein lipase (LPL) is a central enzyme in lipoprotein metabolism and is in part responsible for adipocyte lipid accumulation. Catecholamines are known to decrease the activity of LPL in adipocytes, and we have previously demonstrated that this inhibition occurs posttranscriptionally, with a prominent inhibition of LPL translation. To better characterize the inhibition of LPL translation, 3T3-L1 cells were differentiated into adipocytes, and exposed to epinephrine. Epinephrine induced a dose-dependent decrease in LPL synthesis using [35S]methionine incorporation, with no change in LPL mRNA levels, demonstrating translational regulation of LPL in this cell line. The poly A-enriched RNA from epinephrine-treated cells was translated well in vitro, and there was no difference in the polysome profiles from control and epinephrine-treated cells, suggesting that epinephrine did not affect mRNA editing, and did not induce an inhibition of translation initiation. To obtain evidence for the presence of an inhibitory factor, a cytoplasmic extract from control, and epinephrine-treated adipocytes was human. When compared to the control cell extract, the epinephrine-treated cell extract sharply inhibited LPL translation in vitro, yet had no effect on the translation of other mRNAs. Epinephrine-treated cells had fourfold more of this inhibitor activity than control cells, and this translation inhibition was partially reversed by heat treatment. To determine what region of the LPL mRNA was involved in the translation inhibition, different LPL constructs were synthesized. The inhibitory effect of the epinephrine-treated cell extract was dependent on the presence of the first 40 nucleotides of the 3' (untranslated region UTR) (nucleotides 1599-1638), whereas deletion of the 5' UTR and other areas of the 3' UTR had no effect on translation inhibition. When a sense RNA strand corresponding to this region was added to the in vitro translation reaction, it restored translation towards normal, suggesting that the sense strand was competing for a transacting binding protein. Thus, epinephrine-treated adipocytes produced a transacting factor, probably a protein, that interacted with a region on the LPL mRNA between nucleotides 1599 and 1638, resulting in an inhibition of translation. These studies add new insight into the hormonal regulation of LPL.

Language of Publication

English

Unique Identifier

96066711

MeSH Heading (Major)

Adipocytes|*EN; Lipoprotein Lipase|*BI; RNA, Messenger|GE/*ME; Translation, Genetic|*DE

MeSH Heading

Adrenergic Agonists|PD; Animal; Base Sequence; Dose-Response Relationship, Drug; Epinephrine|PD; Human; Mice; Molecular Sequence Data; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; 3T3 Cells

Publication Type

JOURNAL ARTICLE

ISSN

0021-9738

Country of Publication

UNITED STATES

Record 13 from database: MEDLINE
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Title

Effects of platelet-derived growth factor on the synthesis of lipoprotein lipase in human monocyte-derived macrophages.

Author

Inaba T; Kawamura M; Gotoda T; Harada K; Shimada M; Ohsuga J; Shimano H; Akanuma Y; Yazaki Y; Yamada N

Address

Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.

Source

Arterioscler Thromb Vasc Biol, 1995 Apr, 15:4, 522-8

Abstract

Lipoprotein lipase (LPL), which is secreted by the two predominant cell types in atherosclerotic plaque, macrophages and smooth muscle cells, may be involved in atherosclerosis by generating atherogenic remnant lipoproteins. We investigated the effects of platelet-derived growth factor (PDGF)-BB on the synthesis of LPL by human monocyte-derived macrophages. These cells were cultured in the presence of PDGF-BB for 8 days, after which the enzyme activity, mass, and mRNA levels of LPL were determined. The effect of PDGF-BB was time-dependent and dose-dependent at concentrations of 1 to 10 ng/mL. At 10 ng/mL PDGF-BB enhanced twofold to 2.3-fold the secretion of LPL, and a pulse-labeling study with [35S]methionine revealed that 10 ng/mL PDGF-BB significantly increased the synthesis of LPL. Northern blotting analysis showed that the LPL mRNA level increased dose dependently in macrophages treated with PDGF-BB, and 10 ng/mL PDGF-BB enhanced twofold the expression of LPL mRNA. The protein kinase C inhibitor staurosporine suppressed the effect of PDGF-BB on LPL activity. These results indicate that PDGF-BB stimulated transcription of the LPL gene in human monocyte-derived macrophages through protein kinase C activation and resulted in an increased synthesis of LPL. Therefore, we hypothesize that the augmented synthesis of LPL by PDGF-BB modulates atherosclerosis by influencing lipoprotein metabolism in the vascular wall.

Language of Publication

English

Unique Identifier

95268989

MeSH Heading (Major)

Lipoprotein Lipase|*BI; Macrophages|DE/*EN; Platelet-Derived Growth Factor|*PD

MeSH Heading

Cells, Cultured; Cytokines|PD; Human; Monocytes|CY; RNA, Messenger|AN; Support, Non-U.S. Gov't

Publication Type

JOURNAL ARTICLE

ISSN

1079-5642

Country of Publication

UNITED STATES

Record 14 from database: MEDLINE
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Title

Cholecystokinin release and biliopancreatic secretion in response to selective perfusion of the duodenal loop with aminoacids in man.

Author

Colombel JF; Sutton A; Chayvialle JA; Modigliani R

Address

INSERM U 290, Hôpital Saint-Lazare, Paris, France.

Source

Gut, 1988 Sep, 29:9, 1158-66

Abstract

The aim of this study was to measure the role of the duodenal loop in biliopancreatic secretion in man by infusing various stimuli at the ampulla of Vater and collecting duodenal contents at the ligament of Treitz, above an occluding balloon. Perfusion at 10 ml/min of a first mixture of aminoacids - phenylalanine (47.2 mmol), methionine (38.2 mmol), tryptophan (11 mmol), valine (61.6 mmol) - increased cholecystokinin (CCK) plasma concentrations and duodenal bile salt output (p less than 0.005) as compared with a control electrolyte solution, but did not change pancreatic enzyme secretion significantly; duodenal infusion of another aminoacid mixture - arginine (32.4 mmol), histidine (14.1 mmol), leucine (36 mmol), isoleucine (21.5 mmol), lysine (31 mmol), threonine (23 mmol) - did not change CCK plasma concentrations, bile salt or pancreatic enzyme output. The respective role of duodenal distension and endogenous CCK was investigated by perfusing the first aminoacid solution and the control solution at 2, 5, and 10 ml/min. Changing the perfusion rate of control solution from 2 to 5 ml/min led to a significant increase (p less than 0.01) in pancreatic secretion with no further increase at 10 ml/min. Bile salt output was not influenced by the perfusion rate of control solution. During the perfusion of the aminoacid solution, despite a stepwise increase in CCK release, the only significant change in pancreatic secretion was an increase of lipase output (p less than 0.05) when the infusion rate was raised from 2 to 5 ml/min.(ABSTRACT TRUNCATED AT 250 WORDS)

Language of Publication

English

Unique Identifier

89065418

MeSH Heading (Major)

Amino Acids|*PD; Bile Acids and Salts|ME/*SE; Cholecystokinin|*BL; Duodenum|*DE; Pancreas|EN/*SE

MeSH Heading

Adult; Chymotrypsin|SE; Human; Lipase|SE; Perfusion; Radioimmunoassay; Stimulation, Chemical; Time Factors

Publication Type

JOURNAL ARTICLE

ISSN

0017-5749

Country of Publication

ENGLAND

CAS Registry/EC Number

EC 3.1.1.3 (Lipase); EC 3.4.21.1 (Chymotrypsin); 0 (Amino Acids); 0 (Bile Acids and Salts); 9011-97-6 (Cholecystokinin)

Record 15 from database: MEDLINE
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Title

Apolipoprotein E gene expression in mouse 3T3-L1 adipocytes and human adipose tissue and its regulation by differentiation and lipid content.

Author

Zechner R; Moser R; Newman TC; Fried SK; Breslow JL

Address

Institute of Medical Biochemistry, University of Graz, Austria.

Source

J Biol Chem, 1991 Jun 5, 266:16, 10583-8

Abstract

Apolipoprotein E (apoE) is an important constituent of plasma lipoproteins and a ligand for several lipoprotein receptors. It is produced mainly in the liver but also in several peripheral tissues like brain, adrenal glands, kidney, and macrophages. Some of these tissues also coexpress lipoprotein lipase (LPL), an important enzyme in the metabolism of lipids and lipoproteins. This suggested a possible coordinate expression of these genes and led us to analyze whether adipocytes, a major source of LPL, could also synthesize apoE. Northern blotting experiments showed that apoE mRNA is found in differentiated mouse 3T3-L1 adipocytes as well as biopsies of human adipose tissue maintained in organ culture but not in undifferentiated 3T3-L1 preadipocytes. [35S]Methionine pulse-labeling experiments revealed that apoE protein is produced in human adipose tissue and differentiated mouse 3T3-L1 adipocytes but not in preadipocytes. In biosynthetic labeling experiments, most apoE was found to be cell associated even after prolonged chase periods. Heparin treatment of the cultured cells did not enhance apoE secretion. During differentiation of 3T3-L1 cells, the onset of apoE gene expression was later than that of LPL. The apoE mRNA and intracellular apoE protein concentrations increased linearly with time of differentiation, at least through day 11, whereas LPL showed highest expression at day 7 and then declined. The increase in apoE mRNA correlated with the cellular lipid content. Inhibition of lipid accumulation in differentiated cells by biotin deprivation decreased apoE expression. Cholesterol-loading experiments suggested that apoE mRNA expression is regulated by the intracellular free cholesterol content of 3T3-L1 adipocytes. In contrast, the LPL mRNA level was not influenced by biotin deprivation or cholesterol loading. Human recombinant tumor necrosis factor, a potent inhibitor of LPL gene transcription, had no effect on adipocyte apoE mRNA levels. Therefore, although apoE and LPL are both expressed in adipocytes in a differentiation-dependent manner, the time course of their expression differs as do their responses to cellular lipid content and tumor necrosis factor. We conclude that these genes are not coordinately regulated in adipocytes.

Language of Publication

English

Unique Identifier

91244838

MeSH Heading (Major)

Adipose Tissue|CY/*ME; Apolipoproteins E|*GE; Gene Expression Regulation|*

MeSH Heading

Animal; Blotting, Northern; Cells, Cultured; Cholesterol|AN; Chromatography, Thin Layer; Human; Methionine|CH; Mice; Recombinant Proteins|ME; RNA|AN; RNA, Messenger|AN; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; Tumor Necrosis Factor|ME

Publication Type

JOURNAL ARTICLE

ISSN

0021-9258

Country of Publication

UNITED STATES

CAS Registry/EC Number

0 (Apolipoproteins E); 0 (Recombinant Proteins); 0 (RNA, Messenger); 0 (Tumor Necrosis Factor); 57-88-5 (Cholesterol); 63231-63-0 (RNA); 7005-18-7 (Methionine)

Record 16 from database: MEDLINE
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Title

Hyperlipoproteinemia type I in a patient with active lipoprotein lipase in adipose tissue and indications of defective transport of the enzyme.

Author

Fager G; Semb H; Enerbäck S; Olivecrona T; Jonasson L; Bengtsson-Olivecrona G; Camejo G; Bjursell G; Bondjers G

Address

Arterial Biology Group, Wallenberg Laboratory for Cardiovascular Research, Göteborg, Sweden.

Source

J Lipid Res, 1990 Jul, 31:7, 1187-97

Abstract

This paper presents a case of typical hyperlipoproteinemia type I in a young woman. Her serum triglycerides varied between 2 and 90 mmol/l and she had substantial amounts of apolipoprotein B-48 in fasting plasma. She had no detectable lipoprotein lipase (LPL) activity in post-heparin plasma (less than 0.2 percent of normal). Southern blot analysis suggested no major defect in her LPL gene and Northern blot analysis of adipose tissue RNA showed normal-sized LPL-mRNA. A 2-h [35S]methionine incorporation experiment with adipose tissue pieces in vitro showed that she produced normal-sized LPL and had LPL catalytic activity in the tissue. The amounts were, however, only 5-10% of control. No detectable LPL radioactivity or catalytic activity was released from patient tissue even in the presence of heparin in the incubations. Immunofluorescent staining of adipose tissue biopsies from the patient showed LPL immunoreactivity only in adipocytes and little or none within the capillaries. Treatment of immunoprecipitated labeled LPL with endoglycosidase H showed that the oligosaccharide chains on her enzyme were of the high-mannose type and not processed as in controls. Taken together the data suggest that the patient synthesizes a relatively normal LPL protein which is core-glycosylated and folded into active enzyme as in normal subjects, but is not effectively transported via the Golgi to the cell surface.

Language of Publication

English

Unique Identifier

90383602

MeSH Heading (Major)

Adipose Tissue|*EN; Hyperlipoproteinemia|*EN; Lipoprotein Lipase|GE/*ME; Lipoprotein Lipase Deficiency, Familial|*EN

MeSH Heading

Adult; Biological Transport; Blotting, Northern; Blotting, Southern; Case Report; Dietary Fats|AD; Electrophoresis, Polyacrylamide Gel; Female; Fluorescent Antibody Technique; Genes; Heparin|PD; Human; Precipitin Tests; Support, Non-U.S. Gov't; Triglycerides|BL

Publication Type

JOURNAL ARTICLE

ISSN

0022-2275

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.34 (Lipoprotein Lipase); 0 (Dietary Fats); 0 (Triglycerides); 9005-49-6 (Heparin)

Record 17 from database: MEDLINE
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Title

Effects of insulin and dexamethasone on lipoprotein lipase in human adipose tissue.

Author

Appel B; Fried SK

Address

Laboratory of Human Behavior and Metabolism, Rockefeller University, New York, New York 10021.

Source

Am J Physiol, 1992 May, 262:5 Pt 1, E695-9

Abstract

The mechanisms by which insulin and glucocorticoids modulate lipoprotein lipase (LPL) synthesis and degradation were examined in human adipose tissue fragments maintained in organ culture. Tissue fragments were cultured for 7 days in serum-free medium supplemented with or without insulin (7 nM) and with or without dexamethasone (30 nM), a synthetic glucocorticoid. Responses of LPL activity to both insulin and dexamethasone were obtained at doses within the physiological range. At a maximal dose, insulin increased heparin-releasable and total LPL activity (approximately 7-fold) by specifically increasing the rate of LPL synthesis (approximately 5-fold) determined by pulse labeling with [35S]methionine and [35S]cysteine and immunoprecipitation. Dexamethasone added in the presence of insulin increased heparin-releasable and total LPL activity approximately 8-fold but did not alter rates of LPL synthesis compared with insulin alone. Pulse-chase studies showed that the rate of LPL degradation was markedly slowed in the presence of dexamethasone plus insulin compared with insulin alone. These data suggest that, in human adipose tissue, insulin is essential for maintaining rates of LPL synthesis and that cortisol may play a key role in regulating human adipose tissue LPL at the posttranslational level by inhibiting the degradation of newly synthesized LPL.

Language of Publication

English

Unique Identifier

92272168

MeSH Heading (Major)

Adipose Tissue|*EN; Dexamethasone|*PD; Insulin|*PD; Lipoprotein Lipase|*ME

MeSH Heading

Human; Osmolar Concentration; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

0002-9513

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.34 (Lipoprotein Lipase); 11061-68-0 (Insulin); 50-02-2 (Dexamethasone)

Record 18 from database: MEDLINE
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Title

Effect of improved diabetes control on the expression of lipoprotein lipase in human adipose tissue.

Author

Simsolo RB; Ong JM; Saffari B; Kern PA

Address

Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, CA 90048.

Source

J Lipid Res, 1992 Jan, 33:1, 89-95

Abstract

Patients with diabetes commonly manifest hypertriglyceridemia along with decreased adipose tissue lipoprotein lipase (LPL) activity, and improved diabetes control tends to reverse these abnormalities. To better understand the mechanism of regulation of LPL in diabetes, 11 diabetic patients (3 type I, 8 type II) were brought under improved glycemic control, and adipose tissue LPL gene expression was assessed by performing paired fat biopsies. Six of the 11 patients attained improved control with insulin, with a decrease in glycohemoglobin (glyc Hgb) from 13.8 +/- 0.9 to 10.4 +/- 0.6%; 5 patients attained improved control with glyburide (glyc Hgb fell from 14.2 +/- 2.4 to 8.8 +/- 0.6%), and together they demonstrated a lowering of serum triglycerides and total cholesterol. No changes were observed in HDL cholesterol. Improved diabetes control resulted in a significant increase in LPL activity in both the heparin-releasable (HR) and extractable (EXT) fractions of adipose tissue, as well as in LPL immunoreactive mass. The change in LPL activity with improved control was variable, and showed a positive correlation with the HDL levels prior to treatment (r = 0.74, P less than 0.02). When adipose tissue was pulse-labeled with [35S]methionine, there was an increase in isotope incorporation into LPL after treatment, indicating an increase in LPL synthetic rate. However, improved diabetes control resulted in no significant change in LPL mRNA levels. Thus, improved glycemic control resulted in an increase in LPL activity which correlated with each patient's basal high density lipoprotein. This increase in LPL activity was accompanied by an increase in LPL immunoreactive mass, and an increase in LPL synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Language of Publication

English

Unique Identifier

92202800

MeSH Heading (Major)

Adipose Tissue|*EN; Diabetes Mellitus|DT/*EN; Insulin|*TU; Lipoprotein Lipase|*ME; Sulfonylurea Compounds|*TU

MeSH Heading

Adult; Aged; Blotting, Northern; Female; Human; Male; Middle Age; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

0022-2275

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.34 (Lipoprotein Lipase); 0 (Sulfonylurea Compounds); 11061-68-0 (Insulin)

Record 19 from database: MEDLINE
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Title

Detection of hormone-sensitive lipase in various tissues. I. Expression of an HSL/bacterial fusion protein and generation of anti-HSL antibodies.

Author

Kraemer FB; Patel S; Saedi MS; Sztalryd C

Address

Department of Veterans Affairs Medical Center, Palo Alto, CA 94304.

Source

J Lipid Res, 1993 Apr, 34:4, 663-71

Abstract

Hormone-sensitive lipase (HSL) is an intracellular neutral lipase found in a variety of tissues, but primarily in adipose and steroidogenic tissues, that hydrolyzes triglycerides and cholesteryl esters. In the present studies, a portion of rat HSL cDNA was subcloned into a pET expression system and the resulting recombinant fusion protein was over-expressed in E. coli. The approximately 26 kD HSL/fusion protein was used to generate polyclonal antibodies in rabbits that recognize intact HSL (84 kD) in rat adipose tissue, ovary, adrenal, testis, heart, and lung, as well as in human adipose tissue. In addition, there was an approximately 89 kD protein observed in all rat tissues expressing the 84 kD protein. Unique to testes, there was an immunoreactive protein of approximately 102 kD in sexually immature rats, and additional immunoreactive proteins of approximately 113 kD and approximately 127 kD in sexually mature rats. The anti-HSL/fusion protein antibodies could remove approximately 60-80% of total neutral cholesterol esterase activity from extracts of rat adipose tissue and immunoprecipitated a single 84 kD protein after labeling of adipocytes with [32P]orthophosphate. The incorporation of 32P into the 84 kD HSL protein was stimulated 4-fold by incubation of adipocytes with isoproterenol. The half life of [35S]methionine-labeled HSL was approximately 4 h in normal rat adipocytes. The production of an HSL/fusion protein and generation of antibodies that recognize native HSL should be valuable tools in exploring the mechanisms regulating the expression of HSL activity and the function and localization of its immunoreactive proteins.

Language of Publication

English

Unique Identifier

93267200

MeSH Heading (Major)

Adipose Tissue|*EN; Cholesterol Esterase|*AN/GE/IM; Gene Expression Regulation|*; Recombinant Fusion Proteins|*AN/GE/IM

MeSH Heading

Amino Acid Sequence; Animal; Antibody Formation; Female; Human; Immunoblotting; Male; Molecular Sequence Data; Precipitin Tests; Rats; Rats, Sprague-Dawley; Support, U.S. Gov't, Non-P.H.S.; Support, U.S. Gov't, P.H.S.

Publication Type

JOURNAL ARTICLE

ISSN

0022-2275

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1.13 (Cholesterol Esterase); 0 (Recombinant Fusion Proteins)

Record 20 from database: MEDLINE
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Title

Dissociation between pancreatic enzyme secretory and synthetic dose-responses to cholecystokinin in man.

Author

Boyd EJ; Dunbar J; Clarke G; Wormsley KG

Address

Department of Therapeutics, Ninewells Hospital and Medical School, Dundee, U.K.

Source

Int J Pancreatol, 1986 May, 1:1, 29-37

Abstract

The effect of varying the intensity of pancreatic stimulation on the synthesis of human pancreatic enzymes has not previously been studied. We have measured the secretion and synthesis of pancreatic enzymes in response to either secretin alone (1 CU.kg-1.h-1) or secretion plus increasing doses of cholecystokinin (CCK) (0.25, 0.5 or 1.0 IDU.kg-1.h-1). Enzyme synthesis was measured using the incorporation of 75Se-methionine (0.15 mCi (5.6 kBq).kg-1.h-1) into the trichloracetic acid-insoluble fraction of the duodenal aspirate. Outputs of trypsin, chymotrypsin, lipase and protein showed a bell-shaped dose response to increasing doses of cholecystokinin, with maximal outputs occurring in response to secretin plus cholecystokinin 0.5 IDU.kg-1.h-1. The rate of incorporation of 75Se-methionine increased with increasing doses of cholecystokinin and was maximal in response to secretion plus cholecystokinin 1.0 IDU.kg-1.h-1. There was therefore dissociation between the secretory and synthetic responses to increasing doses of cholecystokinin.

Language of Publication

English

Unique Identifier

88089111

MeSH Heading (Major)

Cholecystokinin|*PD; Pancreas|DE/*EN

MeSH Heading

Dose-Response Relationship, Drug; Human; Support, Non-U.S. Gov't

Publication Type

JOURNAL ARTICLE

ISSN

0169-4197

Country of Publication

NETHERLANDS

CAS Registry/EC Number

9011-97-6 (Cholecystokinin)

HealthGate Document

Record 21 from database: MEDLINE
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Title

Genomic organization, sequence analysis, and chromosomal localization of the human carboxyl ester lipase (CEL) gene and a CEL-like (CELL) gene.

Author

Lidberg U; Nilsson J; Strömberg K; Stenman G; Sahlin P; Enerbäck S; Bjursell G

Address

Department of Molecular Biology, University of Göteborg, Sweden.

Source

Genomics, 1992 Jul, 13:3, 630-40

Abstract

The gene encoding human carboxyl ester lipase (CEL), including 1628 bp of the 5'-flanking region, has been isolated and characterized from two overlapping lambda phage clones. The gene spans 9832 bp and contains 11 exons interrupted by 10 introns. The exons range in size from 88 to 204 bp, except for the last exon, which is 841 bp. A major and a minor transcription initiation site were determined 13 and 7 bp, respectively, upstream of the initiator methionine. The nucleotide sequence is identical with that of the previously reported cDNA, except for the third nucleotide in the 5'-untranslated sequence, a C, which in the cDNA is a T. A TAAATA sequence is present 26 nt upstream from the major CAP site, and within the 5'-flanking region there are several putative transcription factor binding sites. Seven Alu repetitive sequence elements are present in the region analyzed. The organization of the human CEL gene is similar to that of the recently reported rat pancreatic cholesterol esterase gene. The CEL gene was assigned to chromosome 9q34-qter, which confirms the recently reported results of Tayler et al. (1991, Genomics 10: 425-431). A previously unknown gene with a striking homology to the human CEL gene, here called the CEL-like gene (CELL), has also been isolated and characterized, including 1724 bp of the 5'-flanking region. The CELL gene, which most likely is a psuedogene, spans 4846 bp, and due to the absence of a 4.8-kb segment, the CEL gene exons 2-7 are not present in the CELL gene. Despite these differences, the CELL gene is transcribed. We have also assigned the CELL gene to a separate locus at chromosome 9q34-qter.

Language of Publication

English

Unique Identifier

92347858; GENBANK; GENBANK/M94579; GENBANK/M94580; GENBANK/M87672; GENBANK/M87673; GENBANK/M87674; GENBANK/M87675; GENBANK/M87676; GENBANK/M87677; GENBANK/M87678; GENBANK/M87679

MeSH Heading (Major)

Carboxylic Ester Hydrolases|*GE

MeSH Heading

Animal; Base Sequence; Chromosome Mapping; Chromosomes, Human, Pair 9|CHROMOSOMES HUMAN PAIR 09; DNA|GE; Exons; Gene Expression Regulation, Enzymologic; Human; Introns; Molecular Sequence Data; Multigene Family; Rats; Sequence Homology, Nucleic Acid; Support, Non-U.S. Gov't; Transcription, Genetic

Publication Type

JOURNAL ARTICLE

ISSN

0888-7543

Country of Publication

UNITED STATES

CAS Registry/EC Number

EC 3.1.1 (Carboxylic Ester Hydrolases); EC 3.1.1.1 (carboxylesterase); 9007-49-2 (DNA)

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