title: Radiation inactivation target-size analysis of soluble guanylate cyclase.
abstract: The soluble form of guanylate cyclase, which is a heterodimer of two subunits with molecular weights of 82,000 and 70,000, was analyzed by radiation inactivation experiments to determine its functional size. Lyophilized crude extract from rat lung or the purified enzyme were irradiated with different doses from 60Co gamma-rays, and the residual activities were measured in the presence or absence of a potent activator, sodium nitroprusside. The target sizes for the basal activity and for the activity in the presence of sodium nitroprusside were calculated from the decay curve was 77 and 192 kDa, respectively, on the crude enzyme, or as 71 and 163 kDa, respectively, on the purified enzyme. The size for the activatable form of the enzyme was more than twice that of the basal activity and close to the size of the holoenzyme, implying that the enzyme activity must reside on one of the subunits and the activation by sodium nitroprusside requires interaction of both subunits.
authors.affiliation: Department of Clinical Laboratory Medicine, Ehime University School of Medicine, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Saheki
authors.forename: T
authors.initials: T
authors.lastname: Kuno
authors.forename: C
authors.initials: C
authors.lastname: Tanaka
authors.forename: N
authors.initials: N
authors.lastname: Takeuchi
authors.forename: F
authors.initials: F
authors.lastname: Murad
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000187:drug effects / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004789:Enzyme Activation / Q000187:drug effects / Q000528:radiation effects; D006157:Guanosine Monophosphate / Q000032:analysis; D006162:Guanylate Cyclase / Q000302:isolation & purification / Q000378:metabolism* / Q000528:radiation effects; D007700:Kinetics; D008168:Lung / Q000201:enzymology*; D008970:Molecular Weight; D009599:Nitroprusside; D051381:Rats
chemical_list: D009599:Nitroprusside; D006157:Guanosine Monophosphate; D006162:Guanylate Cyclase