Characterization, subsite mapping and N-terminal sequence of miliin, a serine-protease isolated from the latex of Euphorbia milii

Characterization, subsite mapping and N-terminal sequence of miliin, a serine-protease isolated from the latex of Euphorbia milii

Author Moro, Luigi P. Google Scholar
Cabral, Hamilton Google Scholar
Okamoto, Débora Noma Autor UNIFESP Google Scholar
Hirata, Izaura Yoshico Autor UNIFESP Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Bonilla-Rodriguez, Gustavo Orlando Google Scholar
Institution Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Abstract Miliin is a serine protease purified from the latex of Euphorbia milii. This work reports the effect of pH and temperature on the catalytic activity of miliin, using fluorescence resonance energy transfer (FRET) substrates. Miliin displayed the highest activity at pH 9 and 35 degrees C. Subsite mapping shows that subsites S-2 to S-2' prefer uncharged residues. the S-2 subsite prefers hydrophobic aliphatic amino acids (Val, Pro and Ile) and defines the cleavage site. This work is the first one that reports subsite mapping of Euphorbiacea proteases. the N-terminal sequence showed higher similarity (40%) with the serine protease LIM9 isolated from Lilium. the presence of Tyr, Pro and Lys at positions 2, 5 and 10 respectively, were observed for most of the serine proteases used for comparison. the N-terminal sequence has striking differences with those reported previously for milin and eumiliin, other serine proteases isolated from the latex of E. milii. (C) 2013 Elsevier B.V. All rights reserved.
Keywords Euphorbia milii
Miliin
Subsite mapping
Serine protease
Plant protease
FRET substrates
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Date 2013-04-01
Published in Process Biochemistry. Oxford: Elsevier B.V., v. 48, n. 4, p. 633-637, 2013.
ISSN 1359-5113 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 633-637
Origin http://dx.doi.org/10.1016/j.procbio.2013.02.017
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000320413900012
URI http://repositorio.unifesp.br/handle/11600/36110

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