Activity of a peptidase secreted by Phanerochaete chrysosporium depends on lysine to subsite S '(1)

Autor(es)

da Silva, Ronivaldo Rodrigues ; Goncalves de Oliveira, Lilian Caroline ; Juliano, Maria Aparecida ; Juliano, Luiz ; Rosa, Jose C. ; Cabral, Hamilton

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 94, p. 10-pg., 2017-01-01.

Resumo

Peptidases are enzymes that catalyze the rupture of peptide bonds. Catalytic specificity studies of these enzymes have illuminated their modes of action and preferred hydrolysis targets. We describe the biochemical characteristics and catalytic specificity of a lysine-dependent peptidase secreted by the basidiomycete fungus Phanerochaete chrysosporium. We attained 5.7-fold purification of a similar to 23-kDa neutral peptidase using size-exclusion (Sephadex G-50 resin) and ion-exchange (Source 15S resin) chromatography. Using the Fluorescence Resonance Energy Transfer substrate Abz-KLRSSKQ-EDDnp, we detected maximal activity at pH 7.0 and 45-55 degrees C. The peptidase retained similar to 80% of its enzymatic activity for a wide range of conditions (pH 4-9; temperatures up to 50 degrees C for 1 h). The peptidase activity was lowered by the ionic surfactants, sodium dodecyl sulfate and cetyltrimethylammonium bromide; the reducing agent, dithiothreitol; the chaotrope, guanidine; copper (II) ion; and the cysteine peptidase specific inhibitors, iodoacetic acid and N-ethylmaleimide. The peptidase preferred the basic amino acids K and R and high selectivity on S'(1) subsite, exhibiting a condition of lysine-dependence to catalysis on anchoring of this subsite. (C) 2016 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 11/06986-0 - Determinação da especificidade de peptidases isoladas de fungos usando peptídeos FRET como substratos
• Beneficiário: Hamilton Cabral
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 12/24703-8 - Análises proteômicas de fungos filamentosos meso e termofílicos expostos aos fatores físico e químico
• Beneficiário: Hamilton Cabral
• Modalidade de apoio: Auxílio à Pesquisa - Regular