Molecular cloning and deduced amino acid sequences of the -subunits of rat and monkey Nad + -isocitrate dehydrogenases

Coimbra, Joao Carlos; Pinto, Iraja Damiani; Wurdig, Norma Luiza; Do Carmo, Dermeval Aparecido

Biochemical Journal 295(2): 347-350

1993


ISSN/ISBN: 0264-6021
DOI: 10.1042/bj2950347
Accession: 067228520

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Abstract
A 600 bp cDNA fragment encoding part of the gamma-subunit of pig heart NAD(+)-isocitrate dehydrogenase (ICDH gamma) was 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 clones encoding the complete mature forms of the gamma-subunit from rat epididymis and monkey testis cDNA libraries. Comparison of the deduced amino acid sequences of the rat and monkey subunits and the partial sequence of the pig heart enzyme revealed a remarkably high level of sequence identity. The relationship between the deduced amino acid sequences of the NAD(+)-ICDH gamma-subunits and those of nonmammalian NAD(+)- and NADP(+)-ICDH subunits is discussed.