Selective neurotensin-derived internally quenched fluorogenic substrates for neurolysin (EC 3.4.24.16): Comparison with thimet oligopeptidase (EC 3.4.24.15) and neprilysin (EC 3.4.24.11)

Selective neurotensin-derived internally quenched fluorogenic substrates for neurolysin (EC 3.4.24.16): Comparison with thimet oligopeptidase (EC 3.4.24.15) and neprilysin (EC 3.4.24.11)

Autor Oliveira, V Google Scholar
Campos, M. Google Scholar
Hemerly, J. P. Google Scholar
Ferro, E. S. Google Scholar
Camargo, ACM Google Scholar
Juliano, M. A. Google Scholar
Juliano, L. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Universidade de São Paulo (USP)
Inst Butantan
Resumo Internally quenched fluorescent peptides derived from neurotensin (pELYENKPRRPYIL) sequence were synthesized and assayed as substrates for neurolysin (EC 3.4.24.16), thimet oligopeptidase (EC 3.4.24.15 or TOP), and neprilysin (EC 3.4.24.11 or NEP), Abz-LYENKPRRPYILQ-EDDnp (where EDDnp is N-(2,4-dinitrophenyl)ethylenediamine and Abz is ortho-aminobenzoic acid) was derived from neurotensin by the introduction of Q-EDDnp at the C-terminal end of peptide and by the substitution of the pyroglutamic (pE) residue at N-terminus for Abz and a series of shorter peptides was obtained by deletion of amino acids residues from C-terminal, N-terminal, or both sides. Neurolysin and TOP hydrolyzed the substrates at P-Y or Y-I or R-R bonds: depending on the sequence and size of the peptides, while NEP cleaved P-Y or Y-I bonds according to its S'(1) specificity. One of these substrates, Abz-NKPRRPQ-EDDnp was a specific and sensitive substrate for neurolysin (k(cat) = 7.0 s(-l), K(m) = 1.19 muM and k(cat)/K(m) = 5882 mM-(1) . s-(1)), while it was completely resistant to NEP and poorly hydrolyzed by TOP and also by prolyl oligopeptidase (EC 3.4.21.26), Neurolysin concentrations as low as 1 pM were detected using this substrate under our conditions and its analogue Abz-NKPRAPQ-EDDnp was hydrolyzed by neurolysin with k(cat) = 14.03 s(-1), K(m) = 0.82 muM, and k(cat)/K(m) = 17,110 mM(-1) . s(-1), being the best substrate so far described for this peptidase. (C) 2001 Academic Press.
Palavra-chave fluorogenic substrates
neurotensin
neurolysin
thimet oligopeptidase
neprilysin
prolyl oligopeptidase
Idioma Inglês
Data de publicação 2001-05-15
Publicado em Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 292, n. 2, p. 257-265, 2001.
ISSN 0003-2697 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 257-265
Fonte http://dx.doi.org/10.1006/abio.2001.5083
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000168902300012
Endereço permanente http://repositorio.unifesp.br/handle/11600/26555

Exibir registro completo




Arquivo

Arquivo Tamanho Formato Visualização

Não existem arquivos associados a este item.

Este item está nas seguintes coleções

Buscar


Navegar

Minha conta