title: Pharmacological Activities and Hydrolysis by Peptidases of [Phospho-Ser(6)]-Bradykinin (pS(6)-BK).
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. (1)H-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.
authors.affiliation: Department of Biophysics, Escola Paulista de Medicina, Federal University of São Paulo, São Paulo/SP, Brazil.
authors.forename: Diego M
authors.initials: DM
authors.lastname: Assis
authors.affiliation: Department of Biophysics, Escola Paulista de Medicina, Federal University of São Paulo, São Paulo/SP, Brazil. Electronic address: juliano.epm@hotmail.com.
authors.forename: Luiz
authors.initials: L
authors.lastname: Juliano
authors.affiliation: Department of Biophysics, Escola Paulista de Medicina, Federal University of São Paulo, São Paulo/SP, Brazil.
authors.forename: Thaysa
authors.initials: T
authors.lastname: Paschoalin
authors.affiliation: Department of Biochemistry, Biophysics and Medicine, São Paulo Federal University, São Paulo/SP, Brazil.
authors.forename: Maria
authors.initials: M
authors.lastname: Kouyoumdjian
authors.affiliation: Department of Pharmacology, Federal University of Santa Catarina, Florianópolis/SC, Brazil.
authors.forename: Joao B
authors.initials: JB
authors.lastname: Calixto
authors.affiliation: Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte/MG, Brazil.
authors.forename: Robson A S
authors.initials: RA
authors.lastname: Santos
authors.affiliation: Transfusion Medicine Unit, Arcispedale Santa Maria Nuova-IRCCS, Reggio Emilia, Italy.
authors.forename: Thelma A
authors.initials: TA
authors.lastname: Pertinhez
authors.affiliation: UMR INSERM U1100, University of Tours, France.
authors.forename: Francis
authors.initials: F
authors.lastname: Gauthier
authors.affiliation: UMR INSERM U1100, University of Tours, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Moreau
authors.affiliation: Department of Biomedical Sciences, College of Medicine, Florida State University, Tallahassee, FL, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Blaber
authors.affiliation: Department of Biophysics, Escola Paulista de Medicina, Federal University of São Paulo, São Paulo/SP, Brazil. Electronic address: map.juliano@gmail.com.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Juliano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001794:Blood Pressure / Q000187:drug effects / Q000502:physiology; D001920:Bradykinin / Q000031:analogs & derivatives* / Q000235:genetics / Q000494:pharmacology*; D006168:Guinea Pigs; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D051379:Mice; D008969:Molecular Sequence Data; D009924:Organ Culture Techniques; D010447:Peptide Hydrolases / Q000235:genetics / Q000494:pharmacology*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar
chemical_list: D010447:Peptide Hydrolases; D001920:Bradykinin
keywords: Bradykinin (PubChem CID: 439201); Kallikreins; Kinin; Kininogen; Peptidase; Peptides; Protease; Tryptase; [Phospho-Ser(6)]-Bradykinin (PubChem CID: 439201)