Topological and functional analysis of the rat liver carnitine palmitoyltransferase 1 expressed in Saccharomyces cerevisiae

FEBS Letters

Cohen, Isabelle¹  Esser, Victoria²  Kohl, Claude¹  Prip-Buus, Carina¹  McGarry, J.Denis²  Girard, Jean¹ 

[1]`Endocrinologie, métabolisme et développement', CNRS-UPR 1524, 9, rue J. Hetzel, 92190 Meudon-Bellevue, France

[2]Department of Internal Medicine and Biochemistry, University of Texas Southwestern Medical Center, Southwestern Medical School, Dallas, TX, USA

关键词: Carnitine palmitoyltransferase;    Outer mitochondrial membrane;    Topology;    Functional study;    Yeast expression;    L-CPT 1;    liver carnitine palmitoyltransferase 1;    IMM;    inner mitochondrial membrane;    IPTG;    isopropyl-β-d-thiogalactoside;    OMM;    outer mitochondrial membrane;    STI;    soybean trypsin inhibitor;   

DOI  :  10.1016/S0014-5793(98)00584-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The rat liver carnitine palmitoyltransferase 1 (L-CPT 1) expressed in Saccharomyces cerevisiae was correctly inserted into the outer mitochondrial membrane and shared the same folded conformation as the native enzyme found in rat liver mitochondria. Comparison of the biochemical properties of the yeast-expressed L-CPT 1 with those of the native protein revealed the same detergent lability and similar sensitivity to malonyl-CoA inhibition and affinity for carnitine. Normal Michaelis-Menten kinetics towards palmitoyl-CoA were observed when careful experimental conditions were used for the CPT assay. Thus, the expression in S. cerevisiae is a valid model to study the structure-function relationships of L-CPT 1.

【 授权许可】

Unknown