Molecular-cloning and deduced amino-acid-sequences of the alpha-and beta-subunits of mammalian NAD(+)-isocitrate dehydrogenase

B J Nichols, Anthony C F Perry, L Hall, R M Denton

Research output: Contribution to journalArticle (Academic Journal)peer-review

Abstract

A 153 bp fragment of the cDNA encoding the beta-subunit of pig heart NAD(+)-isocitrate dehydrogenase (NAD(+)-ICDH) was specifically amplified by PCR, using redundant oligonucleotide primers based on partial peptide sequence data [Huang and Colman (1990) Biochemistry 29, 8266-8273]. This PCR fragment was then used as a probe to isolate cDNA clones encoding the complete mature form of the beta-subunit from a monkey testis cDNA library. Examination of the deduced amino acid sequence of the monkey subunit and the partial sequence of the pig heart enzyme revealed a high level of sequence conservation. In addition, 3 overlapping fragments of the cDNA for the alpha-subunit of monkey NAD(+)-ICDH were amplified using oligonucleotide primers derived from the cDNA sequence of a subunit of bovine NAD(+)-ICDH (EMBL accession no: U07980). These cDNA fragments allow deduction of the amino acid sequence of the alpha-subunit. Since the gamma-subunit of monkey NAD(+)-ICDH has already been cloned [Nichols, Hall, Ferry and Denton (1993) Biochem. J. 295, 347-350], a deduced amino acid sequence is now available for all three subunits of mammalian NAD(+)-ICDH. Interrelationships between these subunits are discussed and they are compared with the two subunits of yeast NAD(+)-ICDH and Escherichia coli NADP(+)-ICDH.

Original languageEnglish
Pages (from-to)917-922
Number of pages6
JournalBiochemical Journal
Volume310
Publication statusPublished - 15 Sep 1995

Keywords

  • DEPENDENT ISOCITRATE DEHYDROGENASE
  • PIG-HEART
  • CALCIUM-IONS
  • SACCHAROMYCES-CEREVISIAE
  • NUCLEOTIDE-SEQUENCE
  • NAD
  • BINDING
  • GENE
  • MITOCHONDRIA
  • DISRUPTION

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