Expression and substrate specificity of a recombinant cysteine proteinase B of Leishmania braziliensis

Expression and substrate specificity of a recombinant cysteine proteinase B of Leishmania braziliensis

Author Lanfranco, Maria F. Google Scholar
Loayza-Muro, Raul Google Scholar
Clark, Daniel Google Scholar
Nunez, Regina Google Scholar
Zavaleta, Amparo I. Google Scholar
Jimenez, Maribel Google Scholar
Meldal, Morten Google Scholar
Coombs, Graham H. Google Scholar
Mottram, Jeremy C. Google Scholar
Izidoro, Mario Autor UNIFESP Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Arevalo, Jorge Google Scholar
Institution Univ Peruana Cayetano Heredia
Inst Salud Carlos III
Carlsberg Lab
Univ Glasgow
Universidade Federal de São Paulo (UNIFESP)
Abstract The cysteine proteinase B of Leishmania parasites is an important virulence factor. in this study we have expressed, isolated and characterized for the first time a recombinant CPB from Leishmania braziliensis, the causative agent of mucocutaneous leishmaniosis. the mature region of the recombinant CPB shares a high percentage identity with its Leishmania mexicana CPB2.8 (rCPB2.8 Delta CTE) counterpart (76.36%) and has identical amino acid residues at the S(1), catalytic triad and S'(1) subsites. Nevertheless, when the kinetics of substrate hydrolysis was measured using a combinatorial library of internally quenched fluorescent peptides based upon the lead sequence Abz-KLRSSKQ-EDDnp, significant differences were obtained. These results suggest that the differences in substrate utilization observed between the L. mexicana and L braziliensis CPs must be related to amino acid modifications outside the core of the active site cleft. Moreover, a potent inhibitor with Pro at P1 and high affinity for L. braziliensis recombinant CPB showed less affinity to L. mexicana CPB 2.8, which preferred Phe, Leu, and Asn at the same position. (c) 2008 Elsevier B.V. All rights reserved.
Keywords Leishmania braziliensis
recombinant protease
cysteine protease B
enzyme kinetic parameters
fluorogenic peptides
peptide inhibitors
Language English
Sponsor INCO-DC Program
United Nations University/Biotechnology Programme for the Latin America and the Caribbean
Danish National Research Foundation
Swedish Agency for Research Cooperation with Developing Countries
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Grant number INCO-DC Program: ERBIC18CT 970225
Date 2008-10-01
Published in Molecular and Biochemical Parasitology. Amsterdam: Elsevier B.V., v. 161, n. 2, p. 91-100, 2008.
ISSN 0166-6851 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 91-100
Origin http://dx.doi.org/10.1016/j.molbiopara.2008.06.005
Access rights Closed access
Type Article
Web of Science ID WOS:000259339400002
URI http://repositorio.unifesp.br/handle/11600/30944

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