Influence of charge distribution at the active site surface on the substrate specificity of human neutrophil protease 3 and elastase - A kinetic and molecular modeling analysis

Influence of charge distribution at the active site surface on the substrate specificity of human neutrophil protease 3 and elastase - A kinetic and molecular modeling analysis

Author Korkmaz, Brice Google Scholar
Hajjar, Eric Google Scholar
Kalupov, Timofey Google Scholar
Reuter, Nathalie Google Scholar
Brillard-Bourdet, Michele Google Scholar
Moreau, Thierry Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Gauthier, Francis Google Scholar
Institution Fac Med Tours
Univ Tours
Univ Bergen
Universidade Federal de São Paulo (UNIFESP)
Abstract The biological functions of human neutrophil protease 3 ( Pr3) differ from those of neutrophil elastase despite their close structural and functional resemblance. Although both proteases are strongly cationic, their sequences differ mainly in the distribution of charged residues. We have used these differences in electrostatic surface potential in the vicinity of their active site to produce fluorescence resonance energy transfer ( FRET) peptide substrates for investigating individual Pr3 subsites. the specificities of subsites S5 to S3' were investigated both kinetically and by molecular dynamic simulations. Subsites S2, S1', and S2' were the main definers of Pr3 specificity. Combinations of results for each subsite were used to deduce a consensus sequence that was complementary to the extended Pr3 active site and was not recognized by elastase. Similar sequences were identified in natural protein substrates such as NF kappa B and p21 that are specifically cleaved by Pr3. FRET peptides derived from these natural sequences were specifically hydrolyzed by Pr3 with specificity constants k(cat)/K-m in the 10(6) M-1 S-1 range. the consensus Pr3 sequence may also be used to predict cleavage sites within putative protein targets like the proform of interleukin-18, or to develop specific Pr3 peptide-derived inhibitors, because none is available for further studies on the physiopathological function of this protease.
Language English
Date 2007-01-19
Published in Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 282, n. 3, p. 1989-1997, 2007.
ISSN 0021-9258 (Sherpa/Romeo, impact factor)
Publisher Amer Soc Biochemistry Molecular Biology Inc
Extent 1989-1997
Origin http://dx.doi.org/10.1074/jbc.M608700200
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000243451300050
URI http://repositorio.unifesp.br/handle/11600/29462

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