Mode of action of tetrahydrolipstatin: a derivative of the naturally occurring lipase inhibitor lipstatin

Borgström, Bengt

doi:10.1016/0005-2760(88)90260-3

Cited by 253 publications

(134 citation statements)

References 19 publications

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“…This almost 1:1 stoichiometry of the interaction with hPL was in good agreement with the results reported for pPL and the lipases from other species [11,14] an extra carbamoyl group at the N-terminal valine of peptide B (Scheme 2). Such a derivative could easily have been generated as an artifact during the thermolysin digestion in the presence of urea.…”

Section: Isolation and Characterization Of [~H] [T*c]thlmodified Pepsupporting

confidence: 91%

Identification of the active‐site serine in human pancreatic lipase by chemical modification with tetrahydrolipstatin

1992

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A chemical modification approach was used in this study to identify the active site serine residue of human pancreatic lipase. Purified human pancreatic lipase was covalently modified by incubation with ['HI, ["Cjtetrahydrolipstatin (THL). a potent inhibitor of pancreatic lipase. The radiolabeled lipase was digested with thermolysin, and the peptides were separated by HPLC. A single THL-peptidc-adduct was obtained which was identical to that obtained earlier from porcine pancreatic lipase. This pentapeptide with the sequence VIGHS is covalently bound to a THL molecule via the side chain hydroxyl group of the serine unit corresponding to Ser-152 of the lipase. The selective cleavage of the THL-serine bond by mild acid treatment resulted in the formation of the B-lactone Ro 404441 in high yield snd clearly proves that THL is attached via an ester bond and with retention of stereochemistry at all chiral centers to the side chain hydroxyl group of Ser-152 of the lipase. The results obtained for human pancreatic lipase corroborate the inhibition mechanism of THL found on the porcine enzyme, and are in full agreement with the identification of the Ser-lS2...His-263...Asp-176 catalytic triad in the X-ray structure of human pancreatic lipase.Human pancreatic lipase active-site serine; Tetrahydrolipstatin (THL); Structure of lipase-THL adduct

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Section: Isolation and Characterization Of [~H] [T*c]thlmodified Pepsupporting

confidence: 91%

Identification of the active‐site serine in human pancreatic lipase by chemical modification with tetrahydrolipstatin

1992

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“…As was previously found with pancreatic lipase and with gastric lipase (Borgstrom, 1988;Gargouri et al, 1991 ;LuthiPeng and Winkler, 1992) the inhibition of lipoprotein lipase by THL was dependent on amphiphils like fatty acids or deoxycholate. This could be explained by possible conformational changes in the proteins caused by the detergents.…”

Section: Discussionsupporting

confidence: 79%

“…The substance tetrahydrolipstatin (THL) derived from lipstatin produced by Streptomyces toxytricini, appears to be a general inhibitor for mammalian lipases (Hadvary et al, 1988;Borgstrom, 1988;Gargouri et al, 1991). The detailed mechanism of the inhibition has this far only been studied for pancreatic lipase.…”

mentioning

confidence: 99%

“…The inhibition is due to covalent binding of THL to Ser152 which is one of the residues in the catalytic triad of this enzyme (HadvGry et al, 1991 ;. The inhibition is reversible due to hydrolysis of the enzyme-inhibitor complex, as studied with carboxyl ester lipase (Borgstrom, 1988;Stalder et al, 1992). The primary product of the THL hydrolysis, &(N-formyl-L-leucyloxy)-/3-hydroxy acid, is not stable because of its isomerization and lactonization (Stadler et al, 1992).…”

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confidence: 99%

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Interactions of lipoprotein lipase with the active‐site inhibitor tetrahydrolipstatin (Orlistat)^R

Lóokene

Škottová

Olivecrona

1994

European Journal of Biochemistry

121

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Lipoprotein lipase (LPL) was rapidly inactivated by low concentrations of the active-site inhibitor tetrahydrolipstatin (THL). The presence of amphiphils (e.g. long-chain fatty acids) or of lipid water interfaces (lipid emulsions) was required for inhibition to occur. Apolipoprotein CII increased the maximal inactivation rate constant by 1.8-fold in the presence of an emulsion of long-chain triacylglycerols, but had no effect in the presence of an emulsion of tributyrylglycerol. The fully inhibited enzyme had a ratio of THLLPL of nearly 2, indicating that both subunits of the LPL homo-dimer bound THL. The THL-LPL complex was stable below pH 7.5. At higher pH reactivation occurred indicating that THL was slowly turned over by the enzyme. The apparent reactivation rate constant was increased about threefold by the presence of lipidwater interfaces.Sucrose density gradient centrifugation revealed that THL induces tetramerisation of LPL. This aggregation was reversible on reactivation of the inhibited enzyme. Binding to heparin was not affected by THL. In contrast, binding to lipid droplets and to lipoproteins was increased, indicating exposure of hydrophobic regions in the inhibited LPL. It is suggested that THL induces local conformational changes in LPL, which may involve opening of the putative surface lid structure which covers the active-site.

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“…Orlistat (tetrahydrolipstatin) is a potent irreversible inhibitor of gastrointestinal lipases [4,5]. It has been shown to inhibit fat absorption and to reduce body weight, and also to lower plasma cholesterol in animals [6,7].…”

Section: Introduction Methodsmentioning

confidence: 99%

Effect on dietary fat absorption of orlistat, administered at different times relative to meal intake.

Hartmann

Hussain

et al. 1993

Brit J Clinical Pharma

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Orlistat (0) is a potent and selective inhibitor of gastrointestinal lipases. The effect on dietary fat absorption following dosing of 0 at different times relative to meals was investigated in a placebo (P) controlled study in 24 hospitalized healthy males. After a 5-day run-in, to accustom the subjects to a diet of 2400 kcal and 77 g fat per day and to establish baseline faecal fat excretion, subjects received, in four parallel groups of 6, over 8 days three times daily doses of 80 mg 0, P, P (group A) or P, 80 mg 0, P (group B) or P, P, 80 mg 0 (group C) or P, P, P (group D) at mid-meal, 1 h and 2 h after mid-meal respectively. Faeces were collected to measure total fat excretion. The mean (s.d.) of faecal fat in percent of dietary fat, after deduction of pre-treatment faecal fat, was (%) 32.8 (8.1), 34.0 (8.8), 26.9 (4.0) and -1.4 (1.7) in groups A, B, C and D respectively. It was concluded that, within the time period investigated, the pharmacological effect of 0 is not critically dependent on the time of dosing relative to meals.

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Mode of action of tetrahydrolipstatin: a derivative of the naturally occurring lipase inhibitor lipstatin

Cited by 253 publications

References 19 publications

Identification of the active‐site serine in human pancreatic lipase by chemical modification with tetrahydrolipstatin

Identification of the active‐site serine in human pancreatic lipase by chemical modification with tetrahydrolipstatin

Interactions of lipoprotein lipase with the active‐site inhibitor tetrahydrolipstatin (Orlistat)^R

Effect on dietary fat absorption of orlistat, administered at different times relative to meal intake.

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Mode of action of tetrahydrolipstatin: a derivative of the naturally occurring lipase inhibitor lipstatin

Cited by 253 publications

References 19 publications

Identification of the active‐site serine in human pancreatic lipase by chemical modification with tetrahydrolipstatin

Identification of the active‐site serine in human pancreatic lipase by chemical modification with tetrahydrolipstatin

Interactions of lipoprotein lipase with the active‐site inhibitor tetrahydrolipstatin (Orlistat)R

Effect on dietary fat absorption of orlistat, administered at different times relative to meal intake.

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Interactions of lipoprotein lipase with the active‐site inhibitor tetrahydrolipstatin (Orlistat)^R