Catalysis by Isomeric Forms of Covalently Labeled F1-ATPase

  • Jui H. Wang
  • Joe C. Wu
  • Vijay Joshi
  • Hua Chuan
  • Jennifer Cesana

Abstract

When bovine heart mitochondrial F1-ATPase was labeled with low concentrations of NBD-Cl* in the dark in the presence of ATP at pH 7, the NBD-label was almost exclusively attached to a specific tyrosine residue until about one label is covalently attached per F1 molecule (Ferguson et al., 1975a). Cleavage of the labeled protein, isolation and sequencing of the labeled polypeptide and comparison with the known amino acid sequence of the β subunit of the F1 showed that the NBD-label is initially attached to Tyr-β311 (Runswick and Walker, 1983; Andrews et al., 1984a; Sutton and Ferguson, 1985b). Several challenging questions concerning these observations remain unanswered. First, why does the NBD-label inhibit the ATPase? Is it because Tyr-β311 is at the catalytic site so that the label interferes directly with the catalytic mechanism, or because Tyr-β311 is not at the catalytic site but its labeling triggers a long-range protein conformation change which inactivates the enzyme? Second, inasmuch as there are three intrinsically identical β subunits in each F1 molecule, why does the labeling of one β subunit per f1inactivates the whole enzyme?

Keywords

Hydrolysis Urea Catalysis Cysteine Respiration 

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Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Jui H. Wang
    • 1
  • Joe C. Wu
    • 1
  • Vijay Joshi
    • 1
  • Hua Chuan
    • 1
  • Jennifer Cesana
    • 1
  1. 1.Bioenergetics Laboratory, Acheson HallState University of New YorkBuffaloUSA

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