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Molecular cloning of the cDNA of mouse mitochondrial NADP‐dependent isocitrate dehydrogenase and the expression of the gene during lymphocyte activation
Author(s) -
Yang Liangpeng,
Luo Hongyu,
Vinay Patrick,
Wu Jiangping
Publication year - 1996
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19960301)60:3<400::aid-jcb11>3.0.co;2-o
Subject(s) - complementary dna , microbiology and biotechnology , biology , isocitrate dehydrogenase , gene expression , messenger rna , molecular cloning , gene , biochemistry , enzyme
The current report documents the molecular cloning of the mouse mitochondrial NADP‐dependent isocitrate dehydronegase (mNADP‐IDH) cDNA. The cDNA was 1,863 bp in length and contained one open reading frame encoding a 523‐residue polypeptide with a predicted molecular weight of 58 kDa. The cDNA and the deduced amino acid (AA) sequence of the mouse mNADP‐IDH had a high degree of homology with those of porcine, bovine, alfalfa, and yeast. The recombinant mNADP‐IDH expressed in Escherichia coli had active enzymatic function, as well as an expected molecular weight. The heart had the highest constitutive expression of the steady‐state mNADP‐IDH mRNA, followed by the kidney, while the expression of the gene in other tissues was low. The enzymatic activity of different tissues was in agreement with their mNADP‐IDH mRNA levels. The resting lymphocytes had low constitutive expression of the gene, but the steady‐state mRNA could be induced 48 h after mitogen stimulation. At the protein level, the resting lymphocytes had low enzymatic activity of mNADP‐IDH, but the activity was augmented fivefold after mitogen stimulation. The cytosolic NADP‐IDH, on the contrary, remained low or undetectable before and after the mitogen stimulation. Based on our current findings as well as the known roles of the mNADP‐IDH in anabolism and in the isocitrate shuttle, it is conceivable that the mNADP‐IDH is necessary for optimizing proliferation in lymphocytes. © 1996 Wiley‐Liss, Inc.