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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Analysis of peptidase activities of a cathepsin B-like (TcoCBc1) from Trypanosoma congolense

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Author(s):
Oliveira, Lilian C. G. [1] ; Okamoto, Debora N. [1] ; Oliveira, Juliana R. [1] ; Kondo, Marcia Y. [1] ; Gouvea, Iuri E. [1] ; Biteau, Nicolas [2] ; Baltz, Theo [2] ; Murakami, Mario T. [3] ; Juliano, Luiz [1] ; Juliano, Maria A. [1]
Total Authors: 10
Affiliation:
[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Biofis, BR-0444040 Sao Paulo - Brazil
[2] Univ Bordeaux Segalen, CNRS, UMR 5234, F-33000 Bordeaux - France
[3] Ctr Nacl Pesquisas Energia & Mat, Lab Nacl Biociencias, BR-13083970 Campinas, SP - Brazil
Total Affiliations: 3
Document type: Journal article
Source: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1844, n. 7, p. 1260-1267, JUL 2014.
Web of Science Citations: 1
Abstract

The substrate specificity of TcoCBc1 was evaluated using two internally quenched fluorescent peptide libraries with randomized sequences designed to detect carboxydipeptidase (Abz-GXXZXK(Dnp)-OH) and endopeptidase (Abz-GXXZXXQ-EDDnp) activities at acidic and neutral pHs, respectively. All the data obtained with TcoCBc1 were compared with those of human cathepsin B, including the pH profiles of the hydrolytic reactions. The most relevant observation is the preference of TcoCBc1 for substrates with a pair of acidic amino acids at positions P-2 and P-1 for its carboxydipeptidase activity and the well acceptance for E and D at P-1 position for endopeptidase activity. These peculiar preferences for negatively charged groups of TcoCBc1 and its requirements for carboxydipeptidase activity were also observed on Abz labeled analogues of bradykinin (Abz-RPPG(down arrow)FSAFR-OH, Abz-RPPG(down arrow)FS(down arrow)AF-OH, Abz-RPPG(down arrow)DE(down arrow)AF-OH) and angiotensin I (Abz-DR(down arrow)VYIHAFHL-OH), where l indicates the cleavage site. TcoCBc1 was modeled based on the atomic coordinates of the cathepsin B from Trypanosoma brucei and the positively charged environment in TcoCBc1 catalytic site contrasts with the negatively charged environment in human cathepsin B. The preferences of S-1 and S-2 subsites of TcoCBc1 for acidic amino acids have to be taken into consideration for future studies of physiological roles of TcoCBc1 as for instance in apoptotic processes of Trypanosoma congolense. (C) 2014 Elsevier B.V. All rights reserved. (AU)

FAPESP's process: 12/50191-4 - Synthesis, kinetic studies and applications of substrates and inhibitors for proteolytic enzymes
Grantee:Maria Aparecida Juliano
Support Opportunities: Research Projects - Thematic Grants