Interaction of inactivated and active ribulose 1,5-bisphosphate carboxylase/oxygenase of Rhodobacter sphaeroides with nucleotides and the chaperonin 60 (GroEL) protein.
Purified inactivated form I ribulose 1,5-bisphosphate carboxylase/oxygenase (form I RubisCO) of Rhodobacter sphaeroides was activated by ATP and, to some extent, by other adenylates and nucleotides. Reactivation in the presence of ATP occurred by a time-dependent and concentration-dependent process which appeared to be irreversible. The carbamylated form of inactivated form I RubisCO was less susceptible to ATP-mediated reactivation than the uncarbamylated inactivated enzyme. In some cases, ATP analogs could mimic the reactivation process; one analog, adenylyl(beta, gamma-methylene)-diphosphonate, was found to partially block ATP-mediated reactivation but could not block reactivation induced by Mg(II). Concomitant with the recovery of enzymatic activity, the migration of the inactivated form I RubisCO on nondenaturing and sodium dodecyl sulfate gels changed from a pattern that was characteristic of inactivated enzyme to a pattern that was identical to that of the active protein. It was further found that discrete proportions of active enzyme and the chaperonin 60 protein of R. sphaeroides aggregated in the presence of ATP. The form I RubisCO is thus proposed to contain a specific ATP-binding site that may contribute to both the regulation of activity and the assembly of active enzyme.
Wang
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Department of Microbiology, Biotechnology Center, Ohio State University, Columbus 43210.
Tabita
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D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D018834:Chaperonin 60; D004789:Enzyme Activation / Q000187:drug effects; D006360:Heat-Shock Proteins / Q000378:metabolism; D007527:Isoenzymes / Q000187:drug effects; D046911:Macromolecular Substances; D012242:Rhodobacter sphaeroides / Q000201:enzymology*; D012273:Ribulose-Bisphosphate Carboxylase / Q000187:drug effects / Q000378:metabolism*
D001426:Bacterial Proteins; D018834:Chaperonin 60; D006360:Heat-Shock Proteins; D007527:Isoenzymes; D046911:Macromolecular Substances; D000255:Adenosine Triphosphate; D012273:Ribulose-Bisphosphate Carboxylase