Pharmacological Activities and Hydrolysis by Peptidases of [Phospho-Ser(6)]-Bradykinin (pS(6)-BK)

Author(s):Show less -

Assis, Diego M. ^[1] ; Juliano, Luiz ^[1] ; Paschoalin, Thaysa ^[1] ; Kouyoumdjian, Maria ^[2] ; Calixto, Joao B. ^[3] ; Santos, Robson A. S. ^[4] ; Pertinhez, Thelma A. ^[5] ; Gauthier, Francis ^[6] ; Moreau, Thierry ^[6] ; Blaber, Michael ^[7] ; Juliano, Maria A. ^[1]

Affiliation

^[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Biophys, Sao Paulo, SP - Brazil

^[2] Univ Fed Sao Paulo, Dept Biochem Biophys & Med, Sao Paulo, SP - Brazil

^[3] Univ Fed Santa Catarina, Dept Pharmacol, Florianopolis, SC - Brazil

^[4] Univ Fed Minas Gerais, Dept Physiol & Biophys, Belo Horizonte, MG - Brazil

^[5] Arcispedale Santa Maria Nuova IRCCS, Transfus Med Unit, Reggio Emilia - Italy

^[6] Univ Tours, UMR INSERM U1100, F-37041 Tours - France

^[7] Florida State Univ, Coll Med, Dept Biomed Sci, Tallahassee, FL 32306 - USA

• Document type: Journal article
• Source: Biochemical Pharmacology; v. 97, n. 2, p. 203-214, SEP 15 2015.
• Web of Science Citations: 2

Abstract

Phosphorylated kininogen and some of its fragments containing serine phosphorylated bradykinin ({[}pS(6)]-Bk) were identified in human serum and plasma by a phosphoproteomic approach. We report the kininogenase ability of human tissue and plasma kallikreins and tryptase to generate {[}pS(6)]-Bk or Lys-{[}pS(6)]-Bk having as substrate the synthetic human kininogen fluorescent fragment Abz-MISLMKRPPGF {[}pS(386)]PFRSSRI-NH2. The pharmacological assays of {[}pS(6)]-Bk showed it as a full B2 bradykinin receptor agonist in smooth muscle, it produces a portal liver hypertensive response in rat and mouse paw edema that lasts longer than Bk. The rat hypotensive response to infusions of Bk is greater than that of {[}pS(6)]Bk, both if injected through femoral vein or aorta. {[}pS(6)]-Bk was more resistant than Bk to kininase digestion performed with angiotensin converting enzyme, neprilysin, thimet oligopeptidase, aminopeptidase P and carboxypeptidase M. H-1-NMR experiments indicated that {[}pS(6)]-Bk has lower flexibility, with the pS(6)-P-7 bond restricted to the trans conformation, and can explain {[}pS(6)]-Bk resistance to hydrolysis. In conclusion, {[}pS(6)]-Bk presenting lower activity than Bk, with longer lasting effects and being slowly released by kininogenases from synthetic Abz-MISLMKRPPGF{[}pS(386)]PFRSSRI-NH2, suggests that phosphorylation of the kininogens can be an efficient kallikrein-kinin system regulator. (C) 2015 Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 12/50191-4 - Synthesis, kinetic studies and applications of substrates and inhibitors for proteolytic enzymes
• Grantee: Maria Aparecida Juliano
• Support Opportunities: Research Projects - Thematic Grants