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

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

Author Oliveira, Lilian C. G. Autor UNIFESP Google Scholar
Okamoto, Debora N. Autor UNIFESP Google Scholar
Oliveira, Juliana R. Autor UNIFESP Google Scholar
Kondo, Marcia Y. Autor UNIFESP Google Scholar
Gouvea, Iuri E. Autor UNIFESP Google Scholar
Biteau, Nicolas Google Scholar
Baltz, Theo Google Scholar
Murakami, Mario T. Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Bordeaux Segalen
Ctr Nacl Pesquisas Energia & Mat
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.
Keywords Protease
Peptide library
Fluorescent substrate
Carboxydipeptidase
Endopeptidase
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Instituto Nacional de Fluidos Complexos (INCT-FCx-Project)
Grant number FAPESP: 12/50191-4R
CNPq: 471340/2011-1
CNPq: 470388/2010-2
Instituto Nacional de Fluidos Complexos (INCT-FCx-Project): 573560/2008-0
Date 2014-07-01
Published in Biochimica Et Biophysica Acta-proteins and Proteomics. Amsterdam: Elsevier B.V., v. 1844, n. 7, p. 1260-1267, 2014.
ISSN 1570-9639 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 1260-1267
Origin http://dx.doi.org/10.1016/j.bbapap.2014.04.001
Access rights Closed access
Type Article
Web of Science ID WOS:000337211100010
URI http://repositorio.unifesp.br/handle/11600/37933

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