Effect of chemical modification of histidines on the copper-induced oligomerization of jack bean urease (EC 3.5.1.5).
Aggregation of jack bean urease (JBU) is associated with alterations of its biological properties, notably the ureolytic and entomotoxic activities. We investigated the influence of metals on protein oligomerization and biological properties. Besides protein aggregation, Cu(2+) induces inhibition of both ureolytic and insecticidal activities of JBU. Chemical modification of histidine residues in JBU with diethylpyrocarbonate (DEPC) decreases its affinity for Cu(2+) and inhibits oligomerization induced by this metal. Furthermore, this modification protects the insecticidal properties of JBU from being inactivated by Cu(2+). Although DEPC-treated JBU displayed lower ureolytic activity, the modified protein is less susceptible to inhibition by Cu(2+) when compared to native enzyme. Our findings show that Cu(2+) promotes JBU aggregation and differently of other heavy metals studied here, it apparently inhibits the ureolytic activity by inducing protein polymerization along with blockage of sulfhydryl groups.
Follmer
Cristian
C
Graduate Program in Molecular and Cellular Biology, Center of Biotechnology, Universidade Federal Rio Grande do Sul, Porto Alegre, CEP 91.501-970, RS, Brazil. follmer@bioqmed.ufrj.br
Carlini
Célia R
CR
D000818:Animals; D003300:Copper / Q000737:chemistry*; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D007887:Fabaceae / Q000201:enzymology*; D006430:Hemiptera; D007306:Insecticides; D007928:Lethal Dose 50; D008970:Molecular Weight; D011487:Protein Conformation; D016019:Survival Analysis; D014510:Urease / Q000737:chemistry*
D007306:Insecticides; D003300:Copper; D014510:Urease