Characterization of urinary metabolites from four synthetic bradykinin potentiating peptides (BPPs) in mice.
BPPs have been identified in the venom of the Bothrops jararaca snake, or deduced from precursor proteins expressed either in the venom gland or in the brain of the snake. Their potentiating activity on bradykinin (Bk) is assumed to occur through a somatic angiotensin-converting enzyme (sACE) inhibitory mechanism. We have demonstrated that synthetic BPPs show remarkable functional differences, despite their high amino acid sequence similarities. Recently, we demonstrated that BPP-10c, after i.p. administration, was found in its intact form and in the form of a unique metabolite (des-Pro(10) BPP-10c) in mouse urine. Given this finding, we selected a number of BPPs with different structure-activities - BPP-5a (<EKWAP), BPP-7a (<EDGPIPP), BPP-9a (<EWPRPQIPP) and BPP-12b (<EWGRPPGPPIPP) - and studied their stability when exposed to the action of endogenous animal proteolytic enzymes. Here we characterized the BPP metabolites in mouse urine by MALDI-TOF mass spectrometry. Biotransformation results indicated the following: BPP-7a showed a higher resistance to proteolytic cleavage; BPP-5a is metabolized in tripeptides (<EKW); BPP-9a was identified in the intact form and in the form of two metabolites (<EWPRP; <EWPRPQIP); and BPP-12a proved to be very susceptible to hydrolysis by proteolytic enzymes. Thus, the results obtained support the hypothesis that diverse biological functions for each BPP could be mediated by different interactions with alternative targets, and not only by sACE inhibition.
Silva
Carlos A
CA
Center for Applied Toxinology CAT-CEPID, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo, SP 05503-900, Brazil. lescovar@butantangov.br
Ianzer
Danielle A
DA
Portaro
Fernanda C V
FC
Konno
Katsuhiro
K
Faria
Marcella
M
Fernandes
Beatriz L
BL
Camargo
Antonio C M
AC
D000595:Amino Acid Sequence; D000818:Animals; D008297:Male; D051379:Mice; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology / Q000652:urine*; D013329:Structure-Activity Relationship
D009842:Oligopeptides; C006791:bradykinin potentiating factors