Carboxylesterase 3 (EC 3.1.1.1) Is a Major Adipocyte Lipase

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 × 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. 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 × 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. Lipolysis in white adipose tissue (WAT) 1The abbreviations used are: WAT, white adipose tissue; BSA, bovine serum albumin; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid; HSL, hormone-sensitive lipase; LPL, lipoprotein lipase; MS, mass spectrometry; NCEH, neutral cholesteryl ester hydrolase; PNPB, p-nitrophenyl butyrate; TAG, triglyceride; TGH, triacylglycerol hydrolase. exerts a major influence on circulating free fatty acid levels and is an important determinant of energy homeostasis and of WAT mass. In WAT adipocytes, hormone-sensitive lipase (HSL) (EC 3.1.1.3) is an 84-kDa cytoplasmic protein that can cleave fatty acids from the first and third carbons of triacylglycerol (1Fredrikson G. Tornqvist H. Belfrage P. Biochim. Biophys. Acta. 1986; 876: 288-293Crossref PubMed Scopus (157) Google Scholar) and can also hydrolyze cholesteryl esters (2Cook K.G. Yeaman S.J. Stralfors P. Fredrikson G. Belfrage P. Eur. J. Biochem. 1982; 125: 245-249Crossref PubMed Scopus (100) Google Scholar, 3Small C.A. Goodacre J.A. Yeaman S.J. FEBS Lett. 1989; 247: 205-208Crossref PubMed Scopus (73) Google Scholar, 4Khoo J.L. Reue K. Steinberg D. Schotz M.C. J. Lipid Res. 1993; 34: 1969-1974Abstract Full Text PDF PubMed Google Scholar). Until recently, HSL was considered to be the principle or the only mediator of lipolysis in WAT. However, gene-targeted HSL-deficient mice with no detectable HSL show active HSL-independent lipolysis (5Osuga J.-I. Ishabashi S. Oka T. Yagyu H. Tozawa R. Fujimoto A. Shionoiri F. Yahagi N. Kreamer F.B. Tsutsumi O. Yamada N. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 787-792Crossref PubMed Scopus (504) Google Scholar, 6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar). Although HSL –/– mice have a severely blunted response to adrenergic stimulation (6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar), the basal lipolytic rate of isolated adipocytes was somewhat greater in HSL –/– cells than in normal cells (5Osuga J.-I. Ishabashi S. Oka T. Yagyu H. Tozawa R. Fujimoto A. Shionoiri F. Yahagi N. Kreamer F.B. Tsutsumi O. Yamada N. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 787-792Crossref PubMed Scopus (504) Google Scholar, 6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar). In HSL –/– mouse embryonic fibroblasts, the basal lipolytic rate is equal in HSL –/– cells and controls. In homogenates of WAT (5Osuga J.-I. Ishabashi S. Oka T. Yagyu H. Tozawa R. Fujimoto A. Shionoiri F. Yahagi N. Kreamer F.B. Tsutsumi O. Yamada N. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 787-792Crossref PubMed Scopus (504) Google Scholar) and of embryonic fibroblasts from HSL –/– mice (7Okazaki H. Osuga J-I. Tamura Y. Yahagi N. Tomita S. Shioniri F. Iizuka Y. Ohashi K. Harada K. Kimura S. Gotoda T. Shimano H. Yamada N. Ishibashi S. Diabetes. 2002; 51: 3368-3375Crossref PubMed Scopus (101) Google Scholar), total TAG hydrolase (lipase) activity is 40–45% of that of normal mice. Therefore, HSL appears to be responsible for hormone-stimulated lipolysis, but the enzyme(s) mediating basal lipolysis are unknown. We thus attempted to identify non-HSL lipase(s) that may be responsible for HSL-independent lipolysis in WAT. We report that carboxylesterase 3 (triacylglycerol hydrolase; TGH), a protein of the endoplasmic reticulum lumen previously implicated in hepatic very low density lipoprotein synthesis (8Lehner R. Verger R. Biochemistry. 1997; 36: 1861-1868Crossref PubMed Scopus (100) Google Scholar), accounts for a major fraction of non-HSL TAG hydrolase activity in WAT. Chemicals—Oleic acid, triolein, phosphatidylcholine, phosphatidylinositol, fatty acid-free bovine serum albumin, p-nitrophenyl butyrate, CHAPS, and the protease inhibitors antipain, pepstatin, and leupeptin were purchased from Sigma. Affi-Gel 102 (aminoalkyl agarose) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride were purchased from Bio-Rad. Glycerol tri-[9,10-3H]oleate and cholesteryl [1-14C]oleate were from Amersham Biosciences. ECL Western blotting reagents were from Roche Applied Science. Dulbecco's modified Eagle's medium and fetal bovine serum were from Invitrogen. Antibodies—Anti-α-glucosidase II and anti-calnexin monoclonal antibodies were from Stressgen Biotechnologies (Victoria, Canada). Polyclonal guinea pig anti-perilipin antibody was from Research Diagnostics Inc. (Flanders, NJ). Alexa Fluor 488 donkey anti-rabbit IgG, Alexa Fluor 594 goat anti-guinea pig IgG, and ProLong Antifade kit were from Molecular Probes Inc. (Eugene, OR). Mice—For WAT protein isolation, C57BL/6 mice were obtained from Charles River (Montreal, Canada). For comparison between HSL –/– and HSL –/– mice, we used gene-targeted HSL-deficient mice (6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar) and control littermates. Ages were as indicated throughout. Enzyme Assays—The activities of esterase (PNPB hydrolase), lipase (glycerol trioleate hydrolase), and neutral cholesteryl oleate hydrolase (NCEH) were assayed as described (9Holm C. Østerlund T. Methods Mol. Biol. 1999; 109: 109-121PubMed Google Scholar). Briefly, to assay PNPB esterase, 34.8 μl of PNPB were dissolved in 1 ml of acetonitrile. For each assay, 10 μl of PNPB/acetonitrile were added to 990 μl of buffer (0.1 m phosphate and 150 mm NaCl, pH 7.5). Following the addition of 50 μl of sample, tubes were incubated at 37 °C for 10 min. The reaction was terminated by adding 3.25 ml of methanol/chloroform/heptane (10:9:7), vigorously vortexed for 10 s, and then centrifuged at 800 × g for 20 min at 21 °C. The upper phase was collected and afterward subjected to incubation at 42 °C for 3 min. The absorbance of released p-nitrophenol was measured at 400 nm. p-Nitrophenol concentration was calculated using its extinction coefficient of 12,000. For lipase activity, in a 4-ml glass vial, 5.9 mg of triolein, including 50 × 106 cpm of [3H]triolein in toluene, plus a phospholipid mixture (30 μl, 20 mg/ml phosphatidylcholine/phosphatidylinositol, 3:1) were dissolved in chloroform. Solvents were evaporated under a gentle stream of N2. 2 ml of 0.1 m potassium phosphate, pH 7.5, was added and sonicated twice in an ice bath each for 1 min, separated by a 1-min interval. An additional 1.0 ml of 0.1 m potassium phosphate, pH 7.5, was added, and the mixture was sonicated 4 × 30 s on ice with a 30-s interval between sonications. Then 1.0 ml of 20% fatty acid-free bovine serum albumin (BSA) prepared in 0.1 m potassium phosphate (pH 7.5) was added. For each assay, 100 μl of substrate was mixed with 100 μl of sample and incubated for 30 min at 37 °C without shaking. The reaction was terminated by the addition of 3.25 ml methanol/chloroform/heptane (10:9: 7). 1.05 ml of 0.1 m potassium carbonate, 0.1 m boric acid, pH 10.5, and 100 μg of nonlabeled oleic acid as carrier were added, vortexed vigorously for 10 s, and centrifuged at 800 × g for 20 min at 21 °C. The upper 1 ml of the upper fraction was added to 10 ml of EcoLite liquid scintillant (ICN, Costa Mesa, CA) for scintillation counting. For the NCEH assay, 1.17 mg of cholesteryl oleate, including 20 × 106 cpm of [14C]cholesteryl oleate, plus a phospholipid mixture (71.4 μl, including phosphatidylcholine (15 mg/ml) and phosphatidylinositol (5 mg/ml), dissolved in chloroform) were placed in a 4-ml glass vial. Solvents were evaporated under a gentle stream of N2. 2 ml of 0.1 m potassium phosphate, pH 7.5, was added, and after incubation at 37 °C for 10 min it was sonicated twice, each time for 1 min, separated by a 1-min interval. An additional 1.0 ml of 0.1 m potassium phosphate, pH 7.5, was added and incubated at 37 °C for 10 min. The mixture was sonicated for 4 × 30 s on ice with a 30-s interval between sonications. After sonication, 1.0 ml of 20% fatty acid-free BSA prepared in 0.1 m potassium phosphate (pH 7.5) was added. For each assay, 100 μl of substrate was mixed with 100 μl of sample and incubated for 30 min at 37 °C without shaking. Reaction termination, extraction, and scintillation counting were as above. Subcellular Fractionation—Subcellular fractionation was performed as described (10Rickwood D.C. Centrifugation: A Practical Approach. 2nd Ed. IRL Press, Oxford, UK1984: 161-182Google Scholar). Synthesis of Oleic Acid-linked Affi-Gel 102 Chromatography Resin— Oleic acid-linked Affi-Gel 102 resins were synthesized as described (11Hass M.J. Cichowicz D.J. Bailey D.G. Lipids. 1992; 27: 571-576Crossref Scopus (66) Google Scholar). Preparation of Fat Tissue Sample for Chromatography—Following intracardiac catheterization, a perfusion of normal saline at 0.5 ml/min was performed for about 6 min. Fat was collected after perfusing overnight-fasted 5–6-month-old C57BL/6 mice. Samples were stored at –70 °C until use. WAT (10 g) was homogenized in 20 ml of homogenization buffer (0.25 m sucrose, 1 mm EDTA, 1 mm dithioerythritol, 20 2 antipain, 1 pepstatin, pH and then centrifuged at 100,000 × g for 1 at 4 °C. The infranatant and fat cake fractions were The infranatant was to ml in a using a with a Fat activity was from the fat cake fraction with as 10 ml of were added to fat cake and incubated at °C. After at × g for 10 min at 4 the upper phase was and 10 ml of were added to the After incubation for 30 min at tubes were centrifuged at × g for 10 min at 4 °C. The upper phase was and were at for 10 min. After the addition of 10 ml of homogenization were sonicated and then centrifuged at × g at 4 °C for 10 min. The infranatant was The fat cake was After all fractions of in homogenization buffer were The fat cake was a low protein to an oleic acid-linked agarose Oleic Acid-linked ml of infranatant or ml of fat cake were to the gel Amersham at a rate of followed by with buffer 1 (0.25 m in 10 mm phosphate pH 7.5) and buffer 2 m in 10 mm phosphate at a rate of 1.0 were in a of in 10 mm phosphate pH 7.5, with a 1.0 ml/min was 4 were performed at 4 °C. activity was measured in each using PNPB as substrate. activity peaks were to 1 ml in a and then to a protein liquid chromatography were using 10 mm phosphate buffer (pH 7.5) 100 mm and fractions were assayed for carboxylesterase and lipolytic and by SDS-PAGE, followed by Coomassie were excised for of was performed on the as described T. H. K. FEBS Lett. PubMed Scopus Google Scholar). Samples of protein were by a liquid of a liquid chromatography with CA) and an mass were separated by phase liquid chromatography on a A was and was and A rate was used to the The was the mass protein were on an mass with a or on a mass in the to the were purified and on were the using the protein were by at nm. were using the protein assay kit with BSA as a were under PubMed Scopus Google Scholar) and with Coomassie Western separated by were a in mm mm pH at 100 for 1 Following the was with and 20 in 20 mm pH 150 mm at 4 °C and then incubated with a of antibody (8Lehner R. Verger R. Biochemistry. 1997; 36: 1861-1868Crossref PubMed Scopus (100) Google Scholar). The was for 10 min with 20 and incubated for 1 with a of goat anti-rabbit in After in the antibody was by the ECL Applied as by the of the was performed using a anti-calnexin antibody in a of fibroblasts were and in Dulbecco's modified Eagle's medium fetal bovine after the cells 0.5 mm and 3 were added to the were following adipocytes were with in for 10 min, with in for 2 min, and then with and antibodies in the of TGH was by incubation with Alexa Fluor 488 donkey anti-rabbit and with Alexa Fluor 594 goat were performed at 37 and were at After were glass using ProLong Antifade were with a confocal and NCEH in Subcellular of in about of total lipase activity was in the cytosol, and about was in the fat cake fractions are felt to to the of the lipid and to the suggesting a in We lipase from two trioleate hydrolase and NCEH activities in subcellular fractions of of of in a and of oleic gel two peaks of PNPB esterase activity were in infranatant and fat cake fractions In fractions, the first was from the on the until elution under (10 mm phosphate (pH 7.5) and m The contained with the and by the 2 of esterase and lipase activities from infranatant and fat cake the activity was to 2 of fractions. 2 of fat cake and of infranatant also low but detectable cholesteryl esterase activity. For the of lipase to NCEH activities was in fat cake 2 and in infranatant in to in WAT Therefore, most NCEH activity was the under of PNPB esterase, and NCEH activities following oleic acid chromatography of infranatant and fat cake After analysis of fractions corresponding to peaks 1 and 2 of infranatant and of the fat cake all protein with Coomassie were excised for In analysis of the identified TGH on An is in 4 for two of the obtained from the In the are by of to The other by Coomassie of infranatant 2 are in by gel permeation we that the esterase activity of infranatant 2 was to fractions in the mass of of from the of infranatant its as Following of infranatant 1 analysis of of an Coomassie identified esterase 1 with 1 also contained a of a identified as TGH In fat cake only two were detectable by SDS-PAGE, at about and By the was revealed to be TGH peptides, The was TGH Western blotting of WAT subcellular fractions the distribution of TGH was with that of the and the lumen TGH was in and fat cake fractions. However, in to and a fraction of TGH was also in the By confocal most TGH was TGH with in the lumen and some with on the lipid and in HSL –/– activity in infranatant and fat cake fractions of WAT from HSL –/– mice was about 20% that of HSL –/– controls. HSL –/– mice had no detectable NCEH activity and no detectable HSL by Western blotting In TGH was detectable in HSL –/– and –/– WAT By functional TGH was identified as a major lipase in WAT. In infranatant and fat the of TGH was on the of of the TGH acid in infranatant and in fat In we also a of of the protein in with that from the TGH acid In other WAT were lipoprotein lipase acid and was For activity, the and activity is detectable G. G. T. J. Biol. Full Text Full Text PDF PubMed Scopus Google Scholar). lipase for activity (pH J. Biol. Full Text PDF PubMed Google Scholar) and be under assay (pH 7.5). is and is in is to be of for of lipid HSL is important in lipolysis but to activity. In the infranatant and fat cake fractions were prepared under and the activities of HSL and of other neutral In peaks 1 and 2 were obtained by under and are to the activities of non-HSL and with the that HSL is the neutral cholesteryl esterase of WAT (6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar, H. Osuga J-I. Tamura Y. Yahagi N. Tomita S. Shioniri F. Iizuka Y. Ohashi K. Harada K. Kimura S. Gotoda T. Shimano H. Yamada N. Ishibashi S. Diabetes. 2002; 51: 3368-3375Crossref PubMed Scopus (101) Google Scholar), is no detectable NCEH activity in peaks 1 and However, to the oleic and in TGH is a fraction of the total lipase activity. Therefore, a major fraction of in vitro lipolytic activity in WAT is to with this, lipase activity in homogenates of HSL –/– WAT (5Osuga J.-I. Ishabashi S. Oka T. Yagyu H. Tozawa R. Fujimoto A. Shionoiri F. Yahagi N. Kreamer F.B. Tsutsumi O. Yamada N. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 787-792Crossref PubMed Scopus (504) Google Scholar, 6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar) and in HSL –/– mouse embryonic following in vitro differentiation to adipocytes (7Okazaki H. Osuga J-I. Tamura Y. Yahagi N. Tomita S. Shioniri F. Iizuka Y. Ohashi K. Harada K. Kimura S. Gotoda T. Shimano H. Yamada N. Ishibashi S. Diabetes. 2002; 51: 3368-3375Crossref PubMed Scopus (101) Google Scholar). To a first the of lipase activity to TGH may be to for most or all non-HSL lipase activity in WAT. the of lipase(s) in WAT. Although the esterase activity of 2 can be to TGH, 1 TGH to its esterase activity. The of esterase activity in 1 is to esterase a carboxylesterase to be in and K. S. R. 9: PubMed Scopus Google Scholar). are in a of and of and are implicated in the of ester or T. Scopus Google Scholar). activity in WAT T. H. N. Biochim. Biophys. Acta. 1982; PubMed Scopus Google Scholar, T. H. J. Biol. 1992; Full Text PDF Google Scholar). To is the first of esterase 1 in WAT. in tissue to be lipase activity is in fat cake 2 and infranatant 2 that TGH is the major non-HSL lipase in fractions. In fat cake only two were TGH and In WAT infranatant TGH is of a of By gel permeation the esterase activity in is to the of of the other of infranatant 2 have or lipase or NCEH is a of density lipoprotein and is in cholesteryl ester thus be to with or other and to influence TAG hydrolase or NCEH However, the substrate for the NCEH and TAG hydrolase are than for Therefore, the levels of are to influence that WAT TGH lipase and NCEH are consistent with in other and other in lipase (8Lehner R. Verger R. Biochemistry. 1997; 36: 1861-1868Crossref PubMed Scopus (100) Google Scholar) and NCEH R. S. Biochem. Biophys. Res. PubMed Scopus Google Scholar) activities were for In HSL –/– WAT we about 20% of the lipase activity in HSL –/– WAT. previously (5Osuga J.-I. Ishabashi S. Oka T. Yagyu H. Tozawa R. Fujimoto A. Shionoiri F. Yahagi N. Kreamer F.B. Tsutsumi O. Yamada N. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 787-792Crossref PubMed Scopus (504) Google Scholar, 6Wang S.P. Laurin N. Himms-Hagen J. Rudnicki M.A. Levy E. Robert M.-F. Pan L. Oligny L. Mitchell G.A. Obes. Res. 2001; 9: 119-128Crossref PubMed Scopus (195) Google Scholar), no detectable NCEH activity was in –/– WAT. By Western TGH was in HSL –/– and –/– WAT. the low activity of TGH cholesteryl the of NCEH activity in WAT be about is with the used in HSL –/– mice have to have of cholesteryl esters in WAT (5Osuga J.-I. Ishabashi S. Oka T. Yagyu H. Tozawa R. Fujimoto A. Shionoiri F. Yahagi N. Kreamer F.B. Tsutsumi O. Yamada N. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 787-792Crossref PubMed Scopus (504) Google Scholar). the of NCEH activity by TGH is in to In vitro activity. In lipolysis is by lipase and by of lipase(s) to of the lipid droplet. for HSL, from to the lipid following a that protein HSL C. Kreamer F.B. Yeaman S.J. J. Biol. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). A is for a protein that the lipid and that may a to by to TAG substrate C. J. Biol. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). that the of TGH a influence in lipolytic in adipocytes. However, the of TGH on lipolysis be from by subcellular fractionation and confocal microscopy that TGH is in the cytosol, in the and also on the lipid TGH previously isolated from pig (8Lehner R. Verger R. Biochemistry. 1997; 36: 1861-1868Crossref PubMed Scopus (100) Google Scholar) and from R. S. Biochem. Biophys. Res. PubMed Scopus Google Scholar). In WAT, we the distribution of TGH with that of the A greater fraction of TGH than of II was in the cytosol, suggesting that the of TGH in may be to be as distribution of and of Although we have no to this, it is that the TGH may be to the TGH, and II were all in fat of may be to of for the lipid subcellular However, on confocal some TGH is in to the lipid and some with that of a of the lipid are consistent with a in between the lipid and previously by of some with the lipid J. Biol. PubMed Scopus Google Scholar) and by that lipid may from the D.J. J. Biochem. Sci. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar, J. Biol. 2001; PubMed Scopus (100) Google Scholar, H. Mol. PubMed Scopus Google Scholar) and of the of other on the lipid H. Mol. PubMed Scopus Google Scholar, S. G. A A. R. E. J. Sci. 2000; PubMed Google Scholar). that TGH, a accounts for most non-HSL lipase activity in WAT at neutral pH in the that may be in lipolysis We for

Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.