Título:	Characterization of JBURE-IIb isoform of Canavalia ensiformis (L.) DC urease.
Autoria:	MULINARI, F. BECKER-RITT, A. B. DEMARTINI, D. R. LIGABUE-BRAUN, R. STANISÇUASKI, F. VERLI, H. FRAGOSO, R. R. SCHROEDER, E. K. CARLINI, C. R. SA, M. F. G. de
Afiliação:	FERNANDA MULINARI, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL ARLETE BEATRIZ BECKER-RITT, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL DIOGO RIBEIRO DEMARTINI, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL RODRIGO LIGABUE-BRAUN, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL FERNANDA STANISÇUASKI, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL HUGO VERLI, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL RODRIGO DA ROCHA FRAGOSO, CPAC EVELYN KOECHE SCHROEDER, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL CÉLIA REGINA CARLINI, UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL MARIA FATIMA GROSSI DE SA, CENARGEN.
Ano de publicação:	2011
Referência:	Biochimica et Biophysica Acta, v. 1814, n. 12, p. 1758-1768, Dec. 2011.
Conteúdo:	Ureases, nickel-dependent enzymes that catalyze the hydrolysis of urea into ammonia and bicarbonate, are widespread in plants, bacteria, and fungi. Previously, we cloned a cDNA encoding a Canavalia ensiformis urease isoform named JBURE-II, corresponding to a putative smaller urease protein (78 kDa) when compared to other plant ureases. Aiming to produce the recombinant protein, we obtained jbure-IIb, with different 3? and 5? ends, encoding a 90 kDa urease. Three peptides unique to the JBURE-II/-IIb protein were detected by mass spectrometry in seed extracts, indicating that jbure-II/-IIb is a functional gene. Comparative modeling indicates that JBURE-IIb urease has an overall shape almost identical to C. ensiformis major urease JBURE-I with all residues critical for urease activity. The cDNA was cloned into the pET101 vector and the recombinant protein was produced in Escherichia coli. The JBURE-IIb protein, although enzymatically inactive presumably due to the absence of Ni atoms in its active site, impaired the growth of a phytopathogenic fungus and showed entomotoxic properties, inhibiting diuresis of Rhodnius prolixus isolated Malpighian tubules, in concentrations similar to those reported for JBURE-I and canatoxin. The antifungal and entomotoxic properties of the recombinant JBURE-IIb apourease are consistent with a protective role of ureases in plants.
Thesagro:	Canavalia Ensiformis
Tipo do material:	Artigo de periódico
Acesso:	openAccess
Aparece nas coleções:	Artigo em periódico indexado (CENARGEN)