Differential effects of polyamine on the cytosolic and mitochondrial NADP-isocitrate dehydrogenases.
Two isozymes of NADP-dependent isocitrate dehydrogenases (EC 1.1.1.42) exist in mammalian tissues: mitochondrial (ICD1) and cytosolic (ICD2). Effects of polyamines such as spermine, spermidine, and putrescine on the cytosolic and mitochondrial NADP-isocitrate dehydrogenases were analyzed kinetically. Spermine activated ICD2, the cytosolic NADP-isocitrate dehydrogenase from rat liver with the increase in the maximal velocity and the decrease in the affinity for the substrates isocitrate and NADP. The activating action of spermine can be explained by "the uncompetitive effect," and the dissociation constant of spermine for the enzyme-substrate complex was determined to be 1.68 mM. Spermidine and putrescine showed little or no effect. ICD1, the mitochondrial form of NADP-isocitrate dehydrogenase from rat and porcine heart was inhibited by spermine effectively, and by spermidine and putrescine to a lesser extent. Spermine inhibited the enzyme competitively with respect to NADP, and noncompetitively with respect to isocitrate. K(i) value of the enzyme for spermine was 1.3 mM. These results suggest that activation by spermine of cytosolic NADP-isocitrate dehydrogenase can enhance the antioxidant activity by regeneration of GSH, and further is responsible for the stimulation of lipid biosynthesis in cytosol. Spermine may contribute to NADPH supply by enhancing transhydrogenase (EC1.6.1.2) activity through the spermine-dependent activation of Ca(2+) -incorporation to mitochondria.
Department of Biochemistry, Aichi Medical University School of Medicine, Yazakokarimata, Nagakute, Aichi, Japan.
Keiko
K
Murakami
Miyako
M
Haneda
Shouko
S
Iwata
Masataka
M
Yoshino
D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects* / Q000201:enzymology; D007521:Isocitrate Dehydrogenase / Q000378:metabolism*; D007700:Kinetics; D050356:Lipid Metabolism; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000201:enzymology; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000201:enzymology; D009249:NADP / Q000378:metabolism; D009250:NADP Transhydrogenases / Q000378:metabolism; D011485:Protein Binding; D011700:Putrescine / Q000494:pharmacology; D051381:Rats; D013095:Spermidine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D013379:Substrate Specificity; D013552:Swine
D013096:Spermine; D009249:NADP; D007521:Isocitrate Dehydrogenase; C039355:isocitrate dehydrogenase (NADP+); D009250:NADP Transhydrogenases; D002118:Calcium; D013095:Spermidine; D011700:Putrescine