Expression of biologically active hormone‐sensitive lipase in mammalian (COS) cells

FEBS Letters

Belfrage, Per¹  Davis, Richard C.²  Schotz, Michael C.²  Fredrikson, Gudrun¹  Holm, Cecilia¹ 

[1]Department of Medical and Physiological Chemistry 4, University of Lund, Lund, Sweden

[2]Research, Veterans Administration, Wadsworth Medical Center, Los Angeles, USA and Department of Medicine, University of California, Los Angeles, USA

关键词: Triacylglycerol lipase;    COS cell;    Expression system;    HSL;    hormone-sensitive lipase;    FLHSL;    full-length HSL cDNA;    F5 HSL;    HSL cDNA lacking the 5′-untranslated region;    LPL;    lipoprotein lipase;    CAT;    chloramphenicol acetyl transferase;    IgG;    immunoglobulin G;    DFP;    diisopropylfluorophosphate;    anti-HSL;    the IgG fraction of a rabbit polyclonal antiserum prepared using rat adipose tissue HSL lipase;    SDS-PAGE;    sodium dodecylsulfate polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(91)80744-N

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

cDNAs encoding rat adipose tissue hormone-sensitive lipase were expressed in COS cells, under the control of the SV40 promoter to half the level in rat adipocytes, the richest native source of the enzyme. A cDNA lacking most of the long 5′-untranslated region of the full-length rat hormone-sensitive lipase cDNA was, with regard to the lipase activity, on the average 70% more efficiently expressed than the full-length cDNA. The recombinant protein was almost identical to hormone-sensitive lipase of rat adipose tissue with respect to specific activity, susceptibility to inhibitors, molecular size, phosphorylation and activation by cyclic AMP-dependent protein kinase. The described eukaryotic expression system will allow analysis of effects of amino acid substitutions introduced into the lipase molecule by site-directed mutagenesis.

【 授权许可】

Unknown