Inhibition of the plasma membrane [H+]-ATPase of Neurospora crassa by N-ethylmaleimide. Protection by nucleotides.
作者： R J Brooker , C W Slayman
DOI: 10.1016/S0021-9258(18)33675-5
关键词:
摘要: Pre-incubation of the plasma membrane [H+]-ATPase Neurospora crassa with sulfhydryl reagent, N-ethylmaleimide (NEM), leads to a marked inhibition ATPase activity. Loss activity depends upon preincubation pH and follows pseudo-first order kinetics respect NEM concentration. MgATP, physiological substrate for activity, protects against inactivation an average dissociation constant 1.5 mM at 0 degrees C. This value agrees well measured Km MgATP hydrolysis (1.3 30 C 0.9 15 C). MgADP also KD 0.18 C; is competitive inhibitor enzyme Ki 0.08 0.09 Free ATP ADP, as other Mg nucleotides (MgGTP, MgCTP, MgUTP) which are hydrolyzed much slower rates than exert smaller protective effects. These results suggest that protect by binding catalytic site ATPase. can therefore be used probe study nature enzyme-ligand interactions.
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