Substrate specificity and inhibition of human kallikrein-related peptidase 3 (KLK3 or PSA) activated with sodium citrate and glycosaminoglycans

Substrate specificity and inhibition of human kallikrein-related peptidase 3 (KLK3 or PSA) activated with sodium citrate and glycosaminoglycans

Author Andrade, Douglas Autor UNIFESP Google Scholar
Assis, Diego M. Autor UNIFESP Google Scholar
Lima, Aurelio Resende Autor UNIFESP Google Scholar
Oliveira, Juliana R. Autor UNIFESP Google Scholar
Araujo, Mariana S. Autor UNIFESP Google Scholar
Blaber, Sachiko I. Google Scholar
Blaber, Michael Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Florida State Univ
Abstract We report the enzymatic properties and substrate specificity of human recombinant KLK3 in the presence of glycosaminoglycans (GAGs) and sodium citrate. This salt is highly concentrated in prostate and in its presence KLK3 had a similar hydrolytic efficiency as chymotrypsin. in contrast to the latter peptidase. KLK3 activated by sodium citrate efficiently hydrolyzed substrates containing R, H and Pat the P1 position. Activated KLK3 also cleaved peptides derived from the bradykinin domain of human kininogen at the same sites as human kallikrein KLK1, but presented low kininogenase activity. Angiotensin I has several sites for hydrolysis by KLK3; however, it was cleaved only at the Y-I bond (DRVY down arrow IHPFHL). Sodium citrate modulated KLK3 conformation as observed by alterations to the intrinsic fluorescence of phenylalanines and tryptophans. Activated KLK3 was reversibly inhibited by Z-Pro-Prolinal and competitively inhibited by ortho-phenantroline. Together, these are noteworthy observations for the future design of specific non-peptide inhibitors of KLK3 and to find natural substrates. (c) 2010 Elsevier Inc. All rights reserved.
Keywords Kininogen
Prostate-specific-antigen
Peptidase
Protease
Kosmotropic salts
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
USPHS/NIH
Grant number USPHS/NIH: 1R15NS057771-01
Date 2010-06-01
Published in Archives of Biochemistry and Biophysics. New York: Elsevier B.V., v. 498, n. 1, p. 74-82, 2010.
ISSN 0003-9861 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 74-82
Origin http://dx.doi.org/10.1016/j.abb.2010.03.022
Access rights Closed access
Type Article
Web of Science ID WOS:000278070300010
URI http://repositorio.unifesp.br/handle/11600/32585

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