Substrate specificity of recombinant cysteine proteinase, CPB, of Leishmania mexicana

Substrate specificity of recombinant cysteine proteinase, CPB, of Leishmania mexicana

Autor Alves, L. C. Google Scholar
Judice, WAS Google Scholar
St Hilaire, P. M. Google Scholar
Meldal, M. Google Scholar
Sanderson, S. J. Google Scholar
Mottram, J. C. Google Scholar
Coombs, G. H. Google Scholar
Juliano, L. Google Scholar
Juliano, M. A. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Carlsberg Lab
Univ Glasgow
Resumo The primary S-1 subsite specificity of a recombinant cysteine, proteinase, CPB2.8 Delta CTE, of Leishmania mexicana was investigated in a systematic way using a series of peptides derived from Abz-KLRFSKQ-EDDnp in which Arg was substituted by all natural amino acids (where Abz is ortho-amino-benzoyl and EDDnp is N-[2,4-dinitrophenyl]-ethylenediamine). the peptides from this series with charged side chain amino acids, Cys, Cys(SBzl), and Thr(OBzl) were well hydrolysed. All other substitutions resulted in peptides that were resistant or hydrolysed very slowly and inhibited the enzyme with Ki values in the range of 9-400 nM. Looking for natural substrates for CPB2.8, we observed that the recombinant enzyme failed to release kinin from human kininogen, an activity earlier observed with cruzipain from Trypanosoma cruzi (Del Nery et al., J. Biol. Chem. 272 (1997) 25713.). This lack of activity seems to be a result of the resistance to hydrolysis of the sequence at the N-terminal site of bradykinin in the human kininogen. the preferences for the S-3, S-2 and S-1' -S-3' for some amino acids were also examined using substrates derived from Abz-KLRFSKQ-EDDnp with variations at Lys, Leu, Phe, Ser and Lys, using the amino acids Ala, Phe, Leu, His or Pro. Peptides with Phe at P-1' presented the highest affinity to the leishmanial enzyme. for comparison, some of the obtained peptides were also assayed with recombinant human cathepsin L and cruzain. the best substrates for CPB2.8 Delta CTE were also well hydrolysed by cathepsin L, however, the best inhibitors of the parasite enzyme have low affinity to cathepsin L. These promising data provide leads for the design of anti-parasitic drugs directed against the leishmanial enzyme. (C) 2001 Elsevier Science B.V. All rights reserved.
Assunto thiol protease
cysteine proteinase
cathepsin L
Idioma Inglês
Data 2001-08-01
Publicado em Molecular and Biochemical Parasitology. Amsterdam: Elsevier B.V., v. 116, n. 1, p. 1-9, 2001.
ISSN 0166-6851 (Sherpa/Romeo, fator de impacto)
Editor Elsevier B.V.
Extensão 1-9
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000170409300001

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