Controlled peptide solvation in portion-mixing libraries of FRET peptides: Improved specificity determination for dengue 2 virus NS2B-NS3 protease and human cathepsin S

Controlled peptide solvation in portion-mixing libraries of FRET peptides: Improved specificity determination for dengue 2 virus NS2B-NS3 protease and human cathepsin S

Author Alves, Fabiana M. Google Scholar
Hirata, Izaura Y. Google Scholar
Gouvea, Iuri E. Google Scholar
Alves, Marcio F. M. Google Scholar
Meldal, Morten Google Scholar
Bromme, Dieter Google Scholar
Juliano, Luiz Google Scholar
Juliano, Maria A. Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Ctr Gamle Carlsberg
Univ British Columbia
Abstract The solubility of peptides in aqueous buffers used for the enzyme assays is a common limitation for all peptide libraries. in principle, the more water-soluble peptides are, the more susceptible they will be to peptidase hydrolysis. We have demonstrated that this bias can be circumvented in a portion-mixing fluorescence resonance energy transfer (FRET) peptide library by introducing k (lysine in the D-form) in both termini of the peptides. This more solvated library and another one without the k were assayed using trypsin and chymotrypsin as standard peptidases with high selectivity for R and K and for hydrophobic F and Y, respectively. Significantly improved consistency of the information on substrate profiles was obtained from the solvated library. the influence of improved solvation on substrate specificity determination was successfully demonstrated by the difference in specificity observed between the two libraries employing the human cathepsin S (accepts acidic, basic, or neutral amino acids at P(1) position) and Dengue 2 virus NS2B-NS3 protease (high specificity to the pair of basic amino acids K-R, R-R, or Q-R/K at P(2)-P(1) positions). in conclusion, hydration of the peptides has a major influence on protease processing, and this bias can be reduced in bound peptide libraries, improving reliability.
Language English
Date 2007-07-01
Published in Journal of Combinatorial Chemistry. Washington: Amer Chemical Soc, v. 9, n. 4, p. 627-634, 2007.
ISSN 1520-4766 (Sherpa/Romeo, impact factor)
Publisher Amer Chemical Soc
Extent 627-634
Origin http://dx.doi.org/10.1021/cc070042k
Access rights Closed access
Type Article
Web of Science ID WOS:000247820100011
URI http://repositorio.unifesp.br/handle/11600/29864

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