Sequence of the bovine 2-oxoglutarate/malate carrier protein: structural relationship to other mitochondrial transport proteins.
The amino acid sequence of the 2-oxoglutarate/malate carrier protein, a component of the inner membranes of mitochondria, has been deduced from the sequences of overlapping cDNA clones. These clones were generated in polymerase chain reactions using, in the first instance, complex mixtures of oligonucleotides as primers and probes, with sequences based upon partial protein sequences of cyanogen bromide fragments of the purified protein. The protein sequence of the carrier, including the initiator methionine, is 314 amino acids long. The mature protein has a modified alpha-amino group, but the nature of this modification and the precise position of the mature N-terminal amino acid have not been ascertained, although it must lie in amino acids 1-4 of the deduced protein sequence. Comparison of the protein sequence with itself and with those of 3 other mitochondrial carrier proteins, ADP/ATP translocase, the phosphate carrier, and the uncoupling protein from brown fat, shows that all 4 proteins contain a 3-fold repeated sequence about 100 amino acids in length, and all the repeats are interrelated. This suggests that the members of this family of proteins have similar structures and mechanisms and that they have evolved from a common origin. The distribution of hydrophobic amino acids in the oxoglutarate/malate carrier supports the view that the domains are folded into similar structural motifs, possibly consisting of two transmembrane alpha-helices joined by an extensive extramembranous hydrophilic region. Clones of cDNA arising from a longer related transcript of the oxoglutarate/malate carrier gene have also been analyzed. They contain 271 additional nucleotides in the 3' noncoding region.(ABSTRACT TRUNCATED AT 250 WORDS)
Runswick
M J
MJ
MRC Laboratory of Molecular Biology, Cambridge, U.K.
Walker
J E
JE
Bisaccia
F
F
Iacobazzi
V
V
Palmieri
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002417:Cattle; D003001:Cloning, Molecular; D007656:Ketoglutaric Acids / Q000378:metabolism*; D008293:Malates / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000378:metabolism*; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010446:Peptide Fragments / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D002352:Carrier Proteins; D007656:Ketoglutaric Acids; D008293:Malates; D024101:Mitochondrial Proteins; D015345:Oligonucleotide Probes; D010446:Peptide Fragments; C066909:2-oxoglutarate-malate carrier protein