Immunological characterization of the mitochondrial 2‐oxoglutarate carrier from liver and heart

Zara, Vincenzo; Benedittis, R. De; Ragan, C I; Palmieri, F.

doi:10.1016/0014-5793(90)81397-7

Cited by 9 publications

(4 citation statements)

References 18 publications

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“…From the results of homozygosity mapping, which identified the locus for Amish microcephaly as 17q25 [Rosenberg et al, 1999], an important early candidate defect for Amish lethal microcephaly was a deficiency of the mitochondrial 2‐ketoglutarate transporter, or OGC, an integral protein of the mitochondrial inner membrane [Bisaccia et al, 1988; Zara et al, 1990]. OGC is a member of a large family of related mitochondrial proteins and serves to transport into mitochondria 2‐ketoglutarate generated extramitochondrially [Palmieri et al, 1996].…”

Section: Discussionmentioning

confidence: 99%

Amish lethal microcephaly: A new metabolic disorder with severe congenital microcephaly and 2‐ketoglutaric aciduria

et al. 2002

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A new metabolic disorder characterized by severe congenital microcephaly, death within the first year, and severe 2‐ketoglutaric aciduria has been found among the Old‐Order Amish of Lancaster County, Pennsylvania. Amish lethal microcephaly segregates as an autosomal recessive disorder and has an unusually high incidence of at least 1 in 500 births. When the infants are well, the urine organic acid profiles show isolated, extreme elevations of 2‐ketoglutaric acid. However, during otherwise simple viral illnesses, the infants often develop a metabolic acidosis, which may follow a lethal course. Cranial magnetic resonance imaging of a single patient showed a smooth, immature brain similar to that of a 20‐week fetus except for a moderate degree of cerebellar vermal hypoplasia. Assay of 2‐ketoglutarate dehydrogenase in cultured lymphoblasts of one patient showed normal activity. Amish lethal microcephaly maps to 17q25 and may be caused by a defect in a mitochondrial inner membrane protein functioning as a 2‐ketoglutarate transporter. © 2002 Wiley‐Liss, Inc.

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Section: Discussionmentioning

confidence: 99%

Amish lethal microcephaly: A new metabolic disorder with severe congenital microcephaly and 2‐ketoglutaric aciduria

et al. 2002

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“…The conjugates corresponding to 100 Mg of coupled peptide in 0.5 mL of PBS were supplemented with 0.5 mL of complete Freund's adjuvant, and male New Zealand white rabbits were immunized as previously described (Boulay et al, 1986). For the generation of the anti-phosphate carrier protein antiserum, 0.2 mg of phosphate carrier purified from bovine heart dissolved in 0.5 mL of 0.9% NaCl was emulsified with an equal volume of complete Freund's adjuvant, and male New Zealand white rabbits were immunized as described by Zara et al (1990). Sera were decomplemented by treatment at 56 °C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Transmembrane topography of the mitochondrial phosphate carrier explored by peptide-specific antibodies and enzymic digestion

Capobianco¹,

Brandolin²,

Palmieri³

1991

Biochemistry

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Two peptides corresponding to the amino acid sequences 1-10 (N-terminal peptide) and 303-313 (C-terminal peptide) of the bovine heart mitochondrial phosphate carrier have been synthesized. After being coupled to ovalbumin, they were injected into rabbits to raise polyclonal antibodies. The specificity of the generated antibodies was tested by enzyme-linked immunosorbent assay (ELISA) and/or Western blot. Anti-N-terminal antibodies and anti-C-terminal antibodies exclusively reacted with the corresponding terminal peptide, they also reacted with the isolated phosphate carrier as well as with the phosphate carrier protein in mitochondrial lysates. Both anti-N-terminal and anti-C-terminal antibodies bound to freeze-thawed mitochondria, indicating that both termini of the membrane-bound phosphate carrier are exposed to the cytoplasmic side of the inner mitochondrial membrane. These immunological data were complemented with results concerning enzymatic cleavage of the membrane-bound phosphate carrier by carboxypeptidase A and by an arginine-specific endoprotease. Carboxypeptidase A markedly decreased the binding of anti-C-terminal antibodies to phosphate carrier in freeze-thawed mitochondria. Arg-endoprotease cleaved the phosphate carrier in inside-out submitochondrial particles, but not in right-side-out particles, yielding two fragments of similar apparent molecular weight (Mr approximately equal to 14.5K), which were immunodetected only by the anti-N-terminal antiserum, and a fragment of Mr approximately equal to 17K which was detected only by the anti-C-terminal antiserum. It appears, therefore, that Arg-endoprotease cleavage sites of the phosphate carrier are present only at the matrix side of the inner mitochondrial membrane, at Arg-140 and/or Arg-152.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…has been detected (Runswick et al, 1987). In the case of the oxoglutarate/malate carrier, antibodies have been raised to the bovine heart and the rat liver proteins (Zara et al, 1990). The cow anti-heart antiserum cross-reacts with the heart proteins in pig, rat, and rabbit, but not with the liver proteins in the same species or in cows.…”

Section: B L Hmentioning

confidence: 99%

Sequence of the bovine 2-oxoglutarate/malate carrier protein: structural relationship to other mitochondrial transport proteins

Runswick¹,

Walker²,

Bisaccia³

et al. 1990

Biochemistry

170

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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)

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Immunological characterization of the mitochondrial 2‐oxoglutarate carrier from liver and heart

Cited by 9 publications

References 18 publications

Amish lethal microcephaly: A new metabolic disorder with severe congenital microcephaly and 2‐ketoglutaric aciduria

Amish lethal microcephaly: A new metabolic disorder with severe congenital microcephaly and 2‐ketoglutaric aciduria

Transmembrane topography of the mitochondrial phosphate carrier explored by peptide-specific antibodies and enzymic digestion

Sequence of the bovine 2-oxoglutarate/malate carrier protein: structural relationship to other mitochondrial transport proteins

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