Mapping of human plasma kallikrein active site by design of peptides based on modifications of a Kazal-type inhibitor reactive site

Mapping of human plasma kallikrein active site by design of peptides based on modifications of a Kazal-type inhibitor reactive site

Author Nunes, Viviane Abreu Autor UNIFESP Google Scholar
Gozzo, Andrezza Justino Autor UNIFESP Google Scholar
Sampaio, Misako Uemura Autor UNIFESP Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Sampaio, Claudio Augusto Machado Autor UNIFESP Google Scholar
Araujo, Mariana da Silva Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Abstract Human plasma kallikrein (huPK) is a proteinase that participates in several biological processes. Although various inhibitors control its activity, members of the Kazal family have not been identified as huPK inhibitors. in order to map the enzyme active site, we synthesized peptides based on the reactive site (PRILSPV) of a natural Kazal-type inhibitor found in Cayman plasma, which is not an huPK inhibitor. As expected, the leader peptide (Abz-SAPRILSPVQ-EDDnp) was not cleaved by huPK. Modifications to the leader peptide at P-1', P-3' and P-4' positions were made according to the sequence of a phage display-generated recombinant Kazal inhibitor (PYTLKWV) that presented huPK-binding ability. Novel peptides were identified as substrates for huPK and related enzymes. Both porcine pancreatic and human plasma kallikreins cleaved peptides at Arg or Lys bonds, whereas human pancreatic kallikrein cleaved bonds involving Arg or a pair of hydrophobic amino acid residues. Peptide hydrolysis by pancreatic kallikrein was not significantly altered by amino acid replacements. the peptide Abz-SAPRILSWVQ-EDDnp was the best substrate and a competitive inhibitor for huPK, indicating that Trp residue at the P-4' position is important for enzyme action.
Keywords kallikreins
proteinase inhibitors
phage display
quenched peptides
Language English
Date 2003-08-01
Published in Journal of Protein Chemistry. New York: Kluwer Academic/plenum Publ, v. 22, n. 6, p. 533-541, 2003.
ISSN 0277-8033 (Sherpa/Romeo, impact factor)
Publisher Kluwer Academic/plenum Publ
Extent 533-541
Origin http://dx.doi.org/10.1023/B:JOPC.0000005503.20628.4e
Access rights Closed access
Type Article
Web of Science ID WOS:000186865800006
URI http://repositorio.unifesp.br/handle/11600/27355

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