Isolation and characterization of Bradykinin potentiating peptides from Agkistrodon bilineatus venom

Abstract

BackgroundSnake venom is a source of many pharmacologically important molecules. Agkistrodon bilineatus commonly known as Cantil, is spread over Central America particularly in Mexico and Costa Rica. From the venom of Agkistrodon bilineatus we have isolated and characterised six hypotensive peptides, and two bradykinin inhibitor peptides. The IC-50 value of four synthesized peptides was studied, towards angiotensin converting enzyme, in order to study the structure-function relationship of these peptides.ResultsThe purification of the peptides was carried out by size exclusion chromatography, followed by reverse phase chromatography. Sequences of all peptides were determined applying MALDI-TOF/TOF mass spectrometry. These hypotensive peptides bear homology to bradykinin potentiating peptides and venom vasodilator peptide. The peptide with m/z 1355.53 (M + H)+1, and the corresponding sequence ZQWAQGRAPHPP, we identified for the first time. A precursor protein containing a fragment of this peptide was reported at genome level, (Uniprot ID P68515), in Bothrops insularis venom gland. These proline rich hypotensive peptides or bradykinin potentiating peptides are usually present in the venom of Crotalinae, and exhibit specificity in binding to the C domain of somatic angiotensin converting enzyme. Four of these hypotensive peptides, were selected and synthesized to obtain the required quantity to study their IC50 values in complex with the angiotensin converting enzyme. The peptide with the sequence ZLWPRPQIPP displayed the lowest IC50 value of 0.64 μM. The IC50 value of the peptide ZQWAQGRAPHPP was 3.63 μM.ConclusionThe canonical snake venom BPPs classically display the IPP motif at the C-terminus. Our data suggest that the replacement of the highly conserved hydrophobic isoleucine by histidine does not affect the inhibitory activity, indicating that isoleucine is not mandatory to inhibit the angiotensin converting enzyme. The evaluation of IC 50 values show that the peptide with basic pI value exhibits a lower IC 50 value.

Snake Venom Extracellular vesicles (SVEVs) reveal wide molecular and functional proteome diversity

“…Proteomic analysis of SVEVs identified the bradykinin inhibitor peptide. Little is known about bradykinin inhibitor peptide and its role to antagonize the bradykinin function during vasodilatation 64 . In this study, the bradykinin inhibitor peptide (TPPAGPDVGPR, Uniprot P0CJ34|BKIP_CROAT) was identified in all SVEVs with high confidence suggesting a conserved role for this biomolecule in the biological function of the SVEVs.…”

Proteomic analysis of human follicular fluid associated with successful in vitro fertilization

“…Ambekar and co-workers carried out SDS-PAGE, OFFGEL and SCX-based separation followed by LC-MS/MS analysis to characterize 480 HFF proteins for a better understanding of folliculogenesis physiology [ 14 ]. Chen and co-workers explored the HFF biomarkers between successfully fertilized oocytes and unfertilized mature oocytes through nano-scale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS), and found 53 peptides to be potential candidates [ 15 ]. Although proteomic researches on HFF deepened our understanding of reproductive process and provided candidates related to oocyte quality, follicle development, IVF outcome and ovarian disorders, it was still essential to fully delineate the HFF networks and pathways involved in the physiology of reproduction and pathophysiology of infertility.…”

Proteomic Analyses of Agkistrodon contortrix contortrix Venom Using 2D Electrophoresis and MS Techniques

“…As with our previous studies [20] two of the peptides are modified by the presence of pyroglutamate residue at the N-terminus, which is typical for the peptides present in the venom [48,49]. All three peptides are responsible for regulating blood pressure and the width of the blood vessel, and two (1063 and 1214 m / z ) have been previously described in other species of Agkistrodon [48]. Two identified bradykinin-potentiating peptides are inhibitors of metalloproteases and act as a hypotensive peptides [48,50].…”

“…All three peptides are responsible for regulating blood pressure and the width of the blood vessel, and two (1063 and 1214 m / z ) have been previously described in other species of Agkistrodon [48]. Two identified bradykinin-potentiating peptides are inhibitors of metalloproteases and act as a hypotensive peptides [48,50]. In turn, the third identified peptide with m / z 1063 is a bradykinin inhibitor peptide, present in many species, clearly evolutionarily conserved [4,51].…”