title: The mitochondrial tricarboxylate carrier.
abstract: The tricarboxylate carrier has recently been purified from rat liver mitochondria by three distinct scientific groups using different methods. A 37-38-kDa protein has been prepared by silca gel 60 chromatography by our group (Claeys and Azzi, 1989; Glerum et al., 1990). The specific citrate transport activity of this preparation is not significantly different from that measured in mitochondria and it is inhibitable by 1,2,3-benzenetricarboxylic acid. Bisaccia et al. (1990) have reported the isolation of a 30-kDa protein by Celite 535 chromatography, and Kaplan's group (Kaplan et al., 1990) have isolated a 32.5-kDa protein by Matrex Orange, Matrex Blue, and Affi-Gel chromatography. Peptide mapping has failed to support any structural homologies between the 37-38-kDa and the 30-32.5-kD proteins. The 38-kD protein is N-terminally blocked. The peptides obtained by several cleavage procedures have been partially sequenced. Their sequence information has been used to obtain different cDNA clones by a dual approach, the polymerase chain reaction and screening of a lambda ZAP cDNA library. The largest cDNA which could be isolated is 2,986 bp in length and contains a 1071-bp-long open reading frame and an unusually long 3' untranslated region, both of which have been completely sequenced. The protein sequence of the carrier from the first in-frame methionine is 322 amino acids in length and exhibits a molecular mass of 35,546. Comparison of the protein sequence to the sequences of the four members of the mitochondrial carrier protein family (ADP/ATP carrier, phosphate carrier, 2-oxoglutarate/malate carrier, and uncoupling protein) does not reveal significant similarity (cf. Walker et al., 1987). A tripartite internal homology, which is a characteristic of these proteins, is not present in the sequence of the tricarboxylate carrier protein. The mRNA for the tricarboxylate carrier is expressed in rat liver and brain, but not in rat heart.
authors.affiliation: Institu für Biochemie und Molekularbiologie, Universität Bern, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Azzi
authors.forename: M
authors.initials: M
authors.lastname: Glerum
authors.forename: R
authors.initials: R
authors.lastname: Koller
authors.forename: W
authors.initials: W
authors.lastname: Mertens
authors.forename: S
authors.initials: S
authors.lastname: Spycher
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D002417:Cattle; D002951:Citrates / Q000378:metabolism; D019343:Citric Acid; D018076:DNA, Complementary / Q000235:genetics; D007425:Intracellular Membranes / Q000737:chemistry; D008930:Mitochondria, Liver / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D051381:Rats
chemical_list: D002352:Carrier Proteins; D002951:Citrates; D018076:DNA, Complementary; C020723:citrate-binding transport protein; D019343:Citric Acid