Involvement of a histidine residue in the interaction between membrane-anchoring protein (QPs) and succinate dehydrogenase in mitochondrial succinate-ubiquinone reductase.
The involvement of a histidine residue of the membrane-anchoring protein (QPs) fraction in reconstitution of succinate dehydrogenase to form succinate-ubiquinone reductase is studied by using a histidine-modifying reagent, diethylpyrocarbonate (DEPC). A maximum inactivation of 80% of reconstitutive activity is obtained when QPs is treated with 1 mM DEPC at 0 degrees C for 30 min in 50 mM Tris-HCl (pH 7.0). DEPC also inactivates about 85% of intact succinate-ubiquinone reductase. The inactivation of succinate-ubiquinone reductase by DEPC is a result of the modification of essential histidine residues of succinate dehydrogenase. The inactivation is not a result of the modification of the histidine residue in QPs which is essential for interaction with succinate dehydrogenase because the QPs dissociated from the inactivated succinate-ubiquinone reductase is active in reconstitution with active succinate-dehydrogenase. Apparently, the essential histidine in QPs is shielded by succinate dehydrogenase and thus inaccessible to DEPC modification in succinate-ubiquinone reductase. The involvement of a histidine residue of QPs in interaction with succinate dehydrogenase is further evident by the presence of 553 nm shoulder on the alpha-absorption peak of reduced cytochrome b-560 (a characteristic of physical association of QPs with succinate dehydrogenase) in the DEPC-inactivated succinate-ubiquinone reductase. This shoulder disappears from a mixture of succinate dehydrogenase and DEPC-treated QPs when reduced with dithionite. About one histidine residue per molecule of QPs is modified in the DEPC-treated sample, suggesting that only one histidine residue is essential for interaction with succinate dehydrogenase. This essential histidine group is located in the smaller subunit (Mr 13,000) of QPs.
Paudel
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HK
Department of Biochemistry, Oklahoma Agriculture Experimental Station, Oklahoma State University, Stillwater 74078.
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Yu
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D004047:Diethyl Pyrocarbonate / Q000378:metabolism / Q000494:pharmacology*; D042963:Electron Transport Complex II; D006639:Histidine; D007700:Kinetics; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013053:Spectrophotometry; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*
D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D006639:Histidine; D010088:Oxidoreductases; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase; D004047:Diethyl Pyrocarbonate