Please use this identifier to cite or link to this item: http://repositorio.unifesp.br/handle/11600/26555
Title: 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)
Authors: Oliveira, V
Campos, M.
Hemerly, J. P.
Ferro, E. S.
Camargo, ACM
Juliano, M. A.
Juliano, L.
Universidade Federal de São Paulo (UNIFESP)
Universidade de São Paulo (USP)
Inst Butantan
Keywords: fluorogenic substrates
neurotensin
neurolysin
thimet oligopeptidase
neprilysin
prolyl oligopeptidase
Issue Date: 15-May-2001
Publisher: Elsevier B.V.
Citation: Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 292, n. 2, p. 257-265, 2001.
Abstract: 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.
URI: http://repositorio.unifesp.br/handle/11600/26555
ISSN: 0003-2697
Other Identifiers: http://dx.doi.org/10.1006/abio.2001.5083
Appears in Collections:Em verificação - Geral

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