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, Perry 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.
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September 1995
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Research Article|
September 15 1995
Molecular cloning and deduced amino acid sequences of the α- and β- subunits of mammalian NAD+-isocitrate dehydrogenase
B J Nichols;
B J Nichols
1Department of Biochemistry, University of Bristol, School of Medical Sciences, Bristol BS8 1TD, U.K.
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A C F Perry;
A C F Perry
1Department of Biochemistry, University of Bristol, School of Medical Sciences, Bristol BS8 1TD, U.K.
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L Hall;
L Hall
1Department of Biochemistry, University of Bristol, School of Medical Sciences, Bristol BS8 1TD, U.K.
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R M Denton
R M Denton
1Department of Biochemistry, University of Bristol, School of Medical Sciences, Bristol BS8 1TD, U.K.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1995 The Biochemical Society, London
1995
Biochem J (1995) 310 (3): 917–922.
Citation
B J Nichols, A C F Perry, L Hall, R M Denton; Molecular cloning and deduced amino acid sequences of the α- and β- subunits of mammalian NAD+-isocitrate dehydrogenase. Biochem J 15 September 1995; 310 (3): 917–922. doi: https://doi.org/10.1042/bj3100917
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