Impact of endosulfan on cytoplasmic and mitochondrial liver malate dehydrogenase from the freshwater catfish (Clarias batrachus).
The impact of a sublethal concentration of an organochlorine pesticide endosulfan on the activity, specific activity, electrophoretic patterns and kinetic properties of crude and purified forms of cytoplasmic malate dehydrogenase (cMDH) and mitochondrial malate dehydrogenase (mMDH) was evaluated in the liver of the freshwater catfish, Clarias batrachus. The endosulfan reduced significantly the activity and the specific activity of cMDH and mMDH, but had no effect on total cytoplasmic and mitochondrial protein contents. The inhibition produced by endosulfan was of mixed non-competitive-uncompetitive type (KiE > KiES) and of mixed competitive-non-competitive type (KiE < KiES) for crude cMDH and mMDH, respectively. The PAGE shows five distinct isoforms (C1 to C5) of cMDH and two isoforms (M1 and M2) of mMDH. The C5-isoform of liver cMDH is predominant and it corresponds to M2-isoform of mMDH. There are no endosulfan-associated differences in the relative charges of crude cMDH and mMDH as well as their purified isoforms, C5-cMDH and M2-mMDH. The relative molecular weights of the purified isoforms are not affected by endosulfan. The purified C5-cMDH and M2-mMDH of endosulfan-treated liver in vivo showed simple non-competitive (KiE = KiES) type of inhibition; whereas in vitro it was of uncompetitive (KiES) and mixed competitive-non-competitive (Ki < KiES) type for the two respective isoforms. G-1-P acts as an uncompetitive (KiES) inhibitor of C5-cMDH and mixed competitive-non-competitive (KiE < KiES) inhibitor of M2-mMDH of the control fish. The inhibitory pattern of G-1-P is modulated by endosulfan in case of C5-cMDH; whereas there is no alteration in case of M2-mMDH. Summarizing, it can be stated that endosulfan exerts an inhibitory effect on crude cMDH and mMDH in vivo, and their purified isoforms (C5-cMDH and M2-mMDH) in vivo as well as in vitro. The impact of endosulfan is mediated through enzyme-substrate-endosulfan (ES-END) complexing for cMDH and enzyme-endosulfan (E-END) complexing for mMDH.
Mishra
R
R
Department of Zoology, Banaras Hindu University, Varanasi, India.
Shukla
S P
SP
D000818:Animals; D002397:Catfishes / Q000378:metabolism*; D002474:Cell-Free System; D003593:Cytoplasm / Q000187:drug effects / Q000201:enzymology; D004726:Endosulfan / Q000009:adverse effects* / Q000493:pharmacokinetics; D007527:Isoenzymes / Q000737:chemistry / Q000187:drug effects / Q000302:isolation & purification; D008099:Liver / Q000648:ultrastructure*; D008291:Malate Dehydrogenase / Q000187:drug effects* / Q000378:metabolism; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology*
D007527:Isoenzymes; D008291:Malate Dehydrogenase; D004726:Endosulfan