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Journal ArticleDOI

Direct evidence that cholesterol ester hydrolase from adrenal cortex is the same enzyme as hormone-sensitive lipase from adipose tissue.

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TLDR
It is concluded that the same protein, or two very similar proteins, catalyses the hydrolysis of cholesterol esters in adrenal cortex and lipolysis in adipose tissue.
Abstract
Several properties of the cytosolic cholesterol ester hydrolase from bovine adrenal cortex were investigated and those properties were compared directly with those of the well-characterised hormone-sensitive lipase, the rate-limiting enzyme in adipose tissue lipolysis. Properties examined included: (a) activity against different substrates; (b) susceptibility to inhibition by NaF, Hg2+ ions and diisopropyl fluorophosphonate; (c) subunit molecular weight as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate; (d) ability to serve as a substrate for cyclic AMP-dependent protein kinase; (e) effect of phosphorylation on enzyme activity; and (f) degradation pattern of polypeptides following limited proteolysis. In all respects the two enzymes exhibited essentially identical characteristics. It is therefore concluded that the same protein, or two very similar proteins, catalyses the hydrolysis of cholesterol esters in adrenal cortex and lipolysis in adipose tissue. The implication of this finding is discussed in relation to the hormonal control of steroidogenesis in adrenal cortex and of lipolysis in adipose tissue.

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Journal ArticleDOI

Molecular Mechanisms Regulating Hormone-Sensitive Lipase and Lipolysis

TL;DR: The current review summarizes the recent advances with regard to hormone-sensitive lipase structure and molecular mechanisms involved in regulating its activity and lipolysis in general.
Journal ArticleDOI

Adipose triglyceride lipase and the lipolytic catabolism of cellular fat stores

TL;DR: The structure, function, and regulation of lipolytic enzymes with a special emphasis on ATGL are focused on, which create the "lipolysome," a complex metabolic network that contributes to the control of lipid and energy homeostasis.
Journal ArticleDOI

Hormone-sensitive lipase control of intracellular tri-(di-)acylglycerol and cholesteryl ester hydrolysis

TL;DR: Accumulating evidence has defined important functions for HSL in normal physiology, affecting adipocyte lipolysis, steroidogenesis, spermatogenesis, and perhaps insulin secretion and insulin action; however, direct links between abnormal expression or genetic variations of HSL and human disorders, such as obesity, insulin resistance, type 2 diabetes, and hyperlipidemia, await further clarification.
Journal ArticleDOI

Identification of Novel Phosphorylation Sites in Hormone-sensitive Lipase That Are Phosphorylated in Response to Isoproterenol and Govern Activation Properties in Vitro

TL;DR: This study used site-directed mutagenesis and two-dimensional phosphopeptide mapping to show that phosphorylation sites other than the previously identified Ser-563 are phosphorylated in HSL in response to isoproterenol stimulation of32P-labeled rat adipocytes and that Ser-659 and Ser-660 are the major activity controlling sites in vitro.
Journal ArticleDOI

Tissue distribution of the AMP-activated protein kinase, and lack of activation by cyclic-AMP-dependent protein kinase, studied using a specific and sensitive peptide assay.

TL;DR: Using this approach, the kinase activity is examined in nine different rat tissues, plus a mouse macrophage cell line, and it is found that there is a correlation between tissues expressing significant levels of peptide Kinase activity and those active in the synthesis or storage of lipids.
References
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Journal ArticleDOI

Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis.

TL;DR: A rapid and convenient method for peptide mapping of proteins has been developed that involves partial enzymatic proteolysis in the presence of sodium dodecyl sulfate and analysis of the cleavage products by polyacrylamide gel electrophoresis.
Journal ArticleDOI

Molecular Weight Determination of Protein-Dodecyl Sulfate Complexes by Gel Electrophoresis in a Discontinuous Buffer System

TL;DR: The retardation coefficient is shown both empirically and theoretically to be a uniform function of molecular weight of protein-SDS complexes over specified ranges, providing a rationale for determining molecular weight from plots of the negative logarithm of relative mobility against molecular weight.
Journal ArticleDOI

Hormone-sensitive lipase of rat adipose tissue. Purification and some properties.

TL;DR: Modulation of hormone-sensitive lipase activity by phosphorylation in uitm as well as in the intact fat cell is demonstrated and indicates that this is a significant mechanism for the hormonal short term regulation of adipose tissue lipolysis.
Journal ArticleDOI

Microcentrifuge desalting : A rapid, quantitative method for desalting small amounts of protein

TL;DR: A method for the highly efficient desalting of 50–200 μl volumes of solution containing from 10 ng to 5 mg of protein has been developed and should be especially useful for the removal of unbound ligand from a rapidly dissociating ligand-binding protein complex.
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