Premium
A liver mitochondrial outer membrane (MOM) fatty acid transfer complex
Author(s) -
Lee Kwangwon,
Kerner Janos,
Hoppel Charles L.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.850.3
Subject(s) - chemistry , voltage dependent anion channel , fatty acid , carnitine , biochemistry , monomer , bacterial outer membrane , fatty acid synthase , gene isoform , membrane protein , fatty acid binding protein , membrane , gene , escherichia coli , organic chemistry , polymer
Carnitine palmitoyltransferase‐I (CPT‐I) catalyzes the regulated step in overall mitochondrial fatty acid oxidation. Here we present both a Blue‐Native PAGE (BNE) and an immunological approach to isolate and characterize the MOM CPT‐I and its interacting partners. When solubilized MOM was subjected to BNE, a series of high molecular weight bands greater than the monomeric CPT‐I (88 kDa) were identified. The proteins in these bands were characterized by in‐gel digestion and sequence analysis by LC‐MS/MS (Center for Proteomics and Bioinformatics). An immunocapture system was designed using an antibody mixture of five CPT‐I specific peptide‐derived IgGs. Immunocaptured proteins were subjected to LC‐MS/MS. Using this method, not only the expected CPT‐I, but also LCAS (long‐chain acyl‐CoA synthase), and VDAC (voltage‐dependent anion channel) isoforms were captured and identified. These accompanying proteins also were identified with BNE. Therefore, we conclude that the MOM contains a complex for fatty acid transfer that consists of LCAS, VDAC and CPT‐I. Supported by the NIH grant DK‐066107.