Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/35597
Title: Characterization of angiotensin I-converting enzyme N-domain selectivity using positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides
Authors: Bersanetti, Patricia A. [UNIFESP]
Sabatini, Regiane A. [UNIFESP]
Matos, Beatriz S. [UNIFESP]
Douglas, Ross G.
Nchinda, Aloysius
Juliano, Maria A. [UNIFESP]
Pesquero, Joao Bosco [UNIFESP]
Sturrock, Edward D.
Carmona, Adriana K. [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
UCT Fac Hlth Sci
Univ Cape Town
Keywords: angiotensin I-converting enzyme
combinatorial libraries
fluorescence resonance energy transfer (FRET) peptides
molecular docking
N-domain selectivity
Issue Date: 1-Dec-2012
Publisher: Walter de Gruyter & Co
Citation: Biological Chemistry. Berlin: Walter de Gruyter & Co, v. 393, n. 12, p. 1547-1554, 2012.
Abstract: Somatic angiotensin I-converting enzyme (ACE) has two homologous active sites (N and C domains) that show differences in various biochemical properties. in a previous study, we described the use of positional-scanning synthetic combinatorial (PS-SC) libraries of fluorescence resonance energy transfer (FRET) peptides to define the ACE C-domain versus N-domain substrate specificity and developed selective substrates for the C-domain (Bersanetti et al., 2004). in the present work, we used the results from the PS-SC libraries to define the N-domain preferences and designed selective substrates for this domain. the peptide Abz-GDDVAK(Dnp)-OH presented the most favorable residues for N-domain selectivity in the P-3 to P-1' positions. the fluorogenic analog Abz-DVAK(Dnp)-OH (Abz=ortho-aminobenzoic acid; Dnp=2,4-dinitrophenyl) showed the highest selectivity for ACE N-domain (k(cat)/K-m = 1.76 mu M-1.s(-1)). Systematic reduction of the peptide length resulted in a tripeptide that was preferentially hydrolyzed by the C-domain. the binding of Abz-DVAK(Dnp)-OH to the active site of ACE N-domain was examined using a combination of conformational analysis and molecular docking. Our results indicated that the binding energies for the N-domain-substrate complexes were lower than those for the C-domain-substrate, suggesting that the former complexes are more stable.
URI: http://repositorio.unifesp.br/handle/11600/35597
ISSN: 1431-6730
Other Identifiers: http://dx.doi.org/10.1515/hsz-2012-0170
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