Purification, cDNA cloning and heterologous expression of the human mitochondrial NADP(+)-dependent malic enzyme

Mitochondrial NADP(+)-dependent malic enzyme (ME; EC 1.1.1.39) has been purified to homogeneity and characterized kinetically from bovine heart. Partial amino acid sequence information allowed amplification of a specific bovine cDNA, which was used to isolate a full-length human cDNA of this isoform...

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Bibliographic Details
Published in:Biochemical journal 1994-12, Vol.304 ( Pt 3) (3), p.687-692
Main Authors: Loeber, G, Maurer-Fogy, I, Schwendenwein, R
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Blotting, Northern
Cattle
Cloning, Molecular
DNA Primers
DNA, Complementary - genetics
DNA, Complementary - isolation & purification
Escherichia coli - enzymology
Escherichia coli - genetics
Female
Hippocampus - enzymology
Humans
Isoenzymes - genetics
Kinetics
Malate Dehydrogenase - genetics
Malate Dehydrogenase - isolation & purification
Malate Dehydrogenase - metabolism
Male
Mitochondria, Heart - enzymology
Molecular Sequence Data
Open Reading Frames
Polymerase Chain Reaction
RNA, Messenger - analysis
Sequence Homology, Amino Acid
Tissue Distribution
Transcription, Genetic
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Summary:Mitochondrial NADP(+)-dependent malic enzyme (ME; EC 1.1.1.39) has been purified to homogeneity and characterized kinetically from bovine heart. Partial amino acid sequence information allowed amplification of a specific bovine cDNA, which was used to isolate a full-length human cDNA of this isoform of ME. The cDNA is 1930 bp long and codes for a protein of 604 amino acids. Comparison of the amino acid sequence of this isoform with published sequences of other human ME isoforms shows stretches of homology interrupted by larger regions with significant differences. The human protein has been expressed in Escherichia coli, and the recombinant human protein has the same kinetic properties as the corresponding protein purified from bovine heart. Northern blot analysis showed a strong tissue-specific transcription with a predominantly high expression-rate in organs with a low division-rate.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj3040687