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PMID:15220344

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Citation

Soni, KG, Lehner, R, Metalnikov, P, O'Donnell, P, Semache, M, Gao, W, Ashman, K, Pshezhetsky, AV and Mitchell, GA (2004) Carboxylesterase 3 (EC 3.1.1.1) is a major adipocyte lipase. J. Biol. Chem. 279:40683-9

Abstract

Hydrolysis of triglycerides is central to energy homeostasis in white adipose tissue (WAT). Hormone-sensitive lipase (HSL) was previously felt to mediate all lipolysis in WAT. Surprisingly, HSL-deficient mice show active HSL-independent lipolysis, suggesting that other lipase(s) also mediate triglyceride hydrolysis. To clarify this, we used functional proteomics to detect non-HSL lipase(s) in mouse WAT. After cell fractionation of intraabdominal WAT, most non-HSL neutral lipase activity is localized in the 100,000 x g infranatant and fat cake fractions. By oleic acid-linked agarose chromatography of infranatant followed by elution in a 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid gradient, we identified two peaks of esterase activity using p-nitrophenyl butyrate as a substrate. One of the peaks contained most of the lipase activity. In the corresponding fractions, gel permeation chromatography and SDS-PAGE, followed by tandem mass spectrometric analysis of excised Coomassie Blue-stained peptides, revealed carboxylesterase 3 (triacylglycerol hydrolase (TGH); EC 3.1.1.1). TGH is also the principle lipase of WAT fat cake extracts. Partially purified WAT TGH had lipase activity as well as lesser but detectable neutral cholesteryl ester hydrolase activity. Western blotting of subcellular fractions of WAT and confocal microscopy of fibroblasts following in vitro adipocytic differentiation are consistent with a distribution of TGH to endoplasmic reticulum, cytosol, and the lipid droplet. TGH is responsible for a major part of non-HSL lipase activity in WAT in vitro and may mediate some or all HSL-independent lipolysis in adipocytes.

Links

PubMed Online version:10.1074/jbc.M400541200

Keywords

Adipocytes/enzymology; Adipocytes/metabolism; Alkanesulfonic Acids/chemistry; Animals; Blotting, Western; Carboxylesterase/metabolism; Cholesterol/chemistry; Chromatography; Chromatography, Agarose; Chromatography, Gel; Cytosol/metabolism; Electrophoresis, Polyacrylamide Gel; Endoplasmic Reticulum/metabolism; Esterases/metabolism; Fibroblasts/metabolism; Hydrolysis; Lipase/metabolism; Lipase/physiology; Lipid Metabolism; Mass Spectrometry; Mice; Mice, Inbred C57BL; Microscopy, Confocal; NIH 3T3 Cells; Oleic Acid/chemistry; Peptides/chemistry; Sterol Esterase/metabolism; Subcellular Fractions/metabolism; Substrate Specificity; Time Factors; Triglycerides/chemistry

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:EST1D

located_in

GO:0005788: endoplasmic reticulum lumen

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:EST1D

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:EST1D

located_in

GO:0005811: lipid droplet

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:EST1D

involved_in

GO:0016042: lipid catabolic process

ECO:0000303: author statement without traceable support used in manual assertion

P

Seeded From UniProt

complete

MOUSE:EST1D

enables

GO:0004771: sterol esterase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:EST1D

enables

GO:0004806: triglyceride lipase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:EST1D

enables

GO:0052689: carboxylic ester hydrolase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:EST1D

involved_in

GO:0046464: acylglycerol catabolic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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