Studies on the subsite specificity of rat nardilysin (N-arginine dibasic convertase)

dc.contributor.authorChow, K. M.
dc.contributor.authorCsuhai, E.
dc.contributor.authorJuliano, Maria Aparecida [UNIFESP]
dc.contributor.authorSt Pyrek, J.
dc.contributor.authorJuliano, Luiz [UNIFESP]
dc.contributor.authorHersh, L. B.
dc.contributor.institutionUniv Kentucky
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionTransylvania Univ
dc.description.abstractThe subsite specificity of rat nardilysin was investigated using fluorogenic substrates of the type 2-amino-benzoyl-GGX(1)X(2)RKX(3)GQ-ethylenediamine-2,4-dinitrophenyl, where P-2, P-2', and P-3 residues were varied. (The nomenclature of Schechter and Berger (Schechter, L, and Berger, A. (1967) Biochem, Biophys, Res. Commun, 27, 157-162) is used where cleavage of a peptide occurs between the P-1 and P-1' residues, and adjacent residues are designated P-2, P-3, P-2', P-3', etc.) There was little effect on II, among different residues at any of these positions, in contrast, residues at each position affected k(cat), with P-2 residues having the greatest effect, the S-3, S-2, and S-2' subsites differed in their amino acid preference, Tryptophan and serine, which produced poor substrates at the P-2 position, were among the best P-2' residues, the specificity at P-3 was generally opposite that of P-2. Residues at P-2, and to a lesser extent at P-3, influenced the cleavage site. At the P-2 position, His, Phe, Tyr,Asn, or Trp produced cleavage at the amino side of the first basic residue. in contrast, a P-2 Ile or Val produced cleavage between the dibasic pair, Other residues produced intermediate effects, the pH dependence for substrate binding showed that the enzyme prefers to bind a protonated histidine, A comparison of the effect of arginine or lysine at the P-1' or P-1 position showed that there is a tendency to cleave on the amino side of arginine and that this cleavage produces the highest k(cat) values.en
dc.description.affiliationUniv Kentucky, Dept Biochem, Lexington, KY 40563 USA
dc.description.affiliationUniv Kentucky, Mass Spectrometry Facil, Lexington, KY 40563 USA
dc.description.affiliationEscola Paulista Med, Dept Biophys, BR-04034 São Paulo, Brazil
dc.description.affiliationTransylvania Univ, Dept Chem, Lexington, KY 40508 USA
dc.description.affiliationUnifespEscola Paulista Med, Dept Biophys, BR-04034 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.identifier.citationJournal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 275, n. 26, p. 19545-19551, 2000.
dc.publisherAmer Soc Biochemistry Molecular Biology Inc
dc.relation.ispartofJournal of Biological Chemistry
dc.rightsAcesso aberto
dc.titleStudies on the subsite specificity of rat nardilysin (N-arginine dibasic convertase)en