Substrate specificity of human kallikrein 2 (hK2) as determined by phage display technology.

Détails

ID Serval
serval:BIB_4257F7F0615E
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Substrate specificity of human kallikrein 2 (hK2) as determined by phage display technology.
Périodique
European journal of biochemistry / FEBS
Auteur(s)
Cloutier S.M., Chagas J.R., Mach J.P., Gygi C.M., Leisinger H.J., Deperthes D.
ISSN
0014-2956
Statut éditorial
Publié
Date de publication
2002
Peer-reviewed
Oui
Volume
269
Numéro
11
Pages
2747-54
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't - Publication Status: ppublish
Résumé
Human glandular kallikrein 2 (hK2) is a trypsin-like serine protease expressed predominantly in the prostate epithelium. Recently, hK2 has proven to be a useful marker that can be used in combination with prostate specific antigen for screening and diagnosis of prostate cancer. The cleavage by hK2 of certain substrates in the proteolytic cascade suggest that the kallikrein may be involved in prostate cancer development; however, there has been very little other progress toward its biochemical characterization or elucidation of its true physiological role. In the present work, we adapt phage substrate technology to study the substrate specificity of hK2. A phage-displayed random pentapeptide library with exhaustive diversity was generated and then screened with purified hK2. Phages displaying peptides susceptible to hK2 cleavage were amplified in eight rounds of selection and genes encoding substrates were transferred from the phage to a fluorescent system using cyan fluorescent protein (derived from green fluorescent protein) that enables rapid determination of specificity constants. This study shows that hK2 has a strict preference for Arg in the P1 position, which is further enhanced by a Ser in P'1 position. The scissile bonds identified by phage display substrate selection correspond to those of the natural biological substrates of hK2, which include protein C inhibitor, semenogelins, and fibronectin. Moreover, three new putative hK2 protein substrates, shown elsewhere to be involved in the biology of the cancer, have been identified thus reinforcing the importance of hK2 in prostate cancer development.
Mots-clé
Green Fluorescent Proteins, Humans, Kinetics, Luminescent Proteins, Peptide Library, Substrate Specificity, Tissue Kallikreins
Pubmed
Web of science
Open Access
Oui
Création de la notice
21/01/2008 17:11
Dernière modification de la notice
20/08/2019 14:44
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