title: Topological and functional analysis of the rat liver carnitine palmitoyltransferase 1 expressed in Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Endocrinologie, métabolisme et développement, CNRS-UPR 1524, 9, Meudon-Bellevue, France. pripbuus@infobiogen.fr
authors.forename: C
authors.initials: C
authors.lastname: Prip-Buus
authors.forename: I
authors.initials: I
authors.lastname: Cohen
authors.forename: C
authors.initials: C
authors.lastname: Kohl
authors.forename: V
authors.initials: V
authors.lastname: Esser
authors.forename: J D
authors.initials: JD
authors.lastname: McGarry
authors.forename: J
authors.initials: J
authors.lastname: Girard
mesh_terms: D000818:Animals; D002331:Carnitine / Q000378:metabolism; D002334:Carnitine O-Palmitoyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology*; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D013347:Subcellular Fractions; D013379:Substrate Specificity
chemical_list: D011993:Recombinant Fusion Proteins; D010171:Palmitoyl Coenzyme A; D002334:Carnitine O-Palmitoyltransferase; D002331:Carnitine