title: Mitochondrial carnitine palmitoyltransferase 1a (CPT1a) is part of an outer membrane fatty acid transfer complex.
abstract: CPT1a (carnitine palmitoyltransferase 1a) in the liver mitochondrial outer membrane (MOM) catalyzes the primary regulated step in overall mitochondrial fatty acid oxidation. It has been suggested that the fundamental unit of CPT1a exists as a trimer, which, under native conditions, could form a dimer of the trimers, creating a hexamer channel for acylcarnitine translocation. To examine the state of CPT1a in the MOM, we employed a combined approach of sizing by mass and isolation using an immunological method. Blue native electrophoresis followed by detection with immunoblotting and mass spectrometry identified large molecular mass complexes that contained not only CPT1a but also long chain acyl-CoA synthetase (ACSL) and the voltage-dependent anion channel (VDAC). Immunoprecipitation with antisera against the proteins revealed a strong interaction between the three proteins. Immobilized CPT1a-specific antibodies immunocaptured not only CPT1a but also ACSL and VDAC, further strengthening findings with blue native electrophoresis and immunoprecipitation. This study shows strong protein-protein interaction between CPT1a, ACSL, and VDAC. We propose that this complex transfers activated fatty acids through the MOM.
authors.affiliation: Center for Mitochondrial Diseases, Cleveland, Ohio 44106-4981, USA.
authors.forename: Kwangwon
authors.initials: K
authors.lastname: Lee
authors.forename: Janos
authors.initials: J
authors.lastname: Kerner
authors.forename: Charles L
authors.initials: CL
authors.lastname: Hoppel
mesh_terms: D000818:Animals; D001692:Biological Transport; D002334:Carnitine O-Palmitoyltransferase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D003066:Coenzyme A Ligases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004586:Electrophoresis; D005227:Fatty Acids / Q000378:metabolism*; D047468:Immunoprecipitation; D008099:Liver / Q000166:cytology; D008297:Male; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism; D051336:Mitochondrial Membranes / Q000201:enzymology / Q000378:metabolism*; D008970:Molecular Weight; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D051381:Rats; D017207:Rats, Sprague-Dawley; D050994:Voltage-Dependent Anion Channels / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D005227:Fatty Acids; D050994:Voltage-Dependent Anion Channels; D002334:Carnitine O-Palmitoyltransferase; D003066:Coenzyme A Ligases