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Peptide agents are regarded as potential candidates for overcoming the life-threatening resistance of pathogenic microorganisms to classic antibiotics. Accordingly, a peptidomic and genomic investigation of natural antimicrobial peptides (AMPs) can provide structure–functional information for designing peptide antibiotics with therapeutic potential. In the present study, we identified nine AMPs from...
A serine protease termed Da-36 was isolated from crude venom of Deinagkistrodon acutus. The enzyme was a single chain protein with an apparent molecular weight of 36,000 on SDS–PAGE with an isoelectric point of 6.59. Da-36 could clot human plasma by cleaving the Aα, Bβ and γ chains of fibrinogen and also exhibited arginine esterase activity. The proteolytic activity of Da-36 toward TAME was strongly...
The requirements to transform a short disintegrin of the RGD clade into an RTS disintegrin, were investigated through the generation of recombinant mutants of ocellatusin in which the RGD tripeptide was substituted for RTS in different positions along the integrin-specificity loop. Any attempt to create an active integrin α 1 β 1 inhibitory motif within the specificity loop of ocellatusin...
Cobra venom factor (CVF) is an anti-complement factor existing in cobra venom. CVF proteins have been purified from the venoms of Naja haje, Naja siamensis, Naja atra, Naja kaouthia, Naja naja, Naja melanoleuca and Austrelaps superbus, but only three full-length cDNA sequences of CVF are available. In the present work, a cobra venom factor termed OVF was purified from the crude venom of Ophiophagus...
Bm-TFF2 is an amphibian trefoil factor purified from the Bombina maxima skin secretion that is highly toxic to mammals. We previously reported that Bm-TFF2 activates human platelets via protease-activated receptor 1. In this study, for a better understanding of platelet activation induced by Bm-TFF2, we used affinity chromatography and pharmacological inhibitors to investigate the downstream signaling...
The king cobra (Ophiophagus hannah) is the largest venomous snake. Despite the components are mainly neurotoxins, the venom contains several proteins affecting blood system. Natriuretic peptide (NP), one of the important components of snake venoms, could cause local vasodilatation and a promoted capillary permeability facilitating a rapid diffusion of other toxins into the prey tissues. Due to the...
Platelet glycoprotein Ib (GPIb) is a primary adhesion receptor and involved in platelet-related disorders. However, it is difficult to study GPIb-specific platelet stimulation using physiological ligands in vivo. GPIb-binding snake C-type lectins (snaclecs) are useful tools for exploring GPIb in vitro because they act on platelets differently. In the present study, a novel GPIb-binding snaclec, named...
Natriuretic peptides (NPs) play crucial roles in human physiology and pathophysiology through natriuresis, dieresis and vasorelaxation. NPs are also one of the important components of snake venoms. However, the low abundance in snake venom hampered the investigation. Here, a novel natriuretic peptide named Na–NP was purified from the cobra Naja atra venom. Na–NP consists of 45 amino acid residues...
Serious clinical symptoms of Trimeresurus jerdonii bite are mainly caused by abnormalities of blood system. We have previously identified and characterized several bioactive components affecting human blood system, such as serine proteases, metalloproteinases and disintegrins. But few snaclec was characterized in the T. jerdonii venom. In this study, a novel snaclec, named jerdonuxin, was isolated,...
Annexin A2 is a unique member of annexin family with multi-functions in membrane physiology, implicated in inflammation and cancer progression. mRNA of Annexin A2 is abundant in the skin of some amphibians. However, no annexin A2 protein has been isolated and characterized from amphibian skin. In this report, a novel annexin A2 protein with apparent molecular weight of 33 kDa and named Bm-ANXA2, was...
Previous in vivo study demonstrated that βγ-CAT, a newly identified non-lens βγ-crystallin and trefoil factor complex from frog Bombina maxima skin secretions, possessed potent lethal toxicity on mammals resulted from hypotension and cardiorespiratory arrest. However, the mechanism of cardiac dysfunction induced by the protein is unknown. Here, we report that βγ-CAT, with dosages of 0.8–3.0nM, elicited...
In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues but their functions are unknown. The molecular mechanisms of trefoil factors, initiators of mucosal healing and being greatly involved in tumorigenesis, have remained elusive. βγ-CAT, which is responsible for the potent hemolytic activity and lethal toxicity on mice of frog Bombina maxima skin secretions, is the first...
In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues, but their functions are unknown. The molecular mechanisms of trefoil factors, initiators of mucosal healing and being greatly involved in tumorigenesis, have remained elusive. βγ-CAT is the first example of a naturally existing multifunctional protein complex of a non-lens βγ-crystallin and a trefoil factor from frog...
An l-amino acid oxidase from Ophiophagus hannah snake venom (Oh-LAAO) was purified by successive gel filtration, ion-exchange and heparin chromatography. Oh-LAAO did not induce platelet aggregation; however, it had potent inhibitory activity on platelet aggregation induced by ADP and U46619, but showed no effect on platelet aggregation induced by thrombin, mucetin, ristocetin and stejnulxin. By RT-PCR...
Serine proteases are widely distributed in viperid snake venoms, but rare in elapid snake venoms. Previously, we have identified a fibrinogenolytic enzyme termed OhS1 from the venom of Ophiophagus hannah. The results indicated that OhS1 might be a serine protease, but there was no structural evidence previously. In the present study, the primary structure of OhS1 was determined by protein sequencing,...
A 50kDa fibrinogenolytic protease, ohagin, from the venom of Ophiophagus hannah was isolated by a combination of gel filtration, ion-exchange and heparin affinity chromatography. Ohagin specifically degraded the alpha-chain of human fibrinogen and the proteolytic activity was completely abolished by EDTA, but not by PMSF, suggesting it is a metalloproteinase. It dose-dependently inhibited platelet...
TSV-DM, a basic metalloproteinase with a molecular weight of 110kDa, was purified from Trimeresurus stejnegeri venom. TSV-DM degraded the Aα chain of fibrinogen more rapidly than the Bβ chain in a dose dependent manner. The cDNA of TSV-DM encoded a polypeptide of 622 amino acid residues, which comprises a signal peptide, proprotein, metalloproteinase domain, spacer, disintegrin-like domain and cysteine-rich...
Hemorrhagic toxins are widely distributed in viperid and crotalid snake venoms. Envenomation of Trimeresurus stejnegeri, a member of Crotalidae family, caused potent systemic and local hemorrhage. Up to now, there is no report on hemorrhage toxins from this venom. In this work, we cloned two cDNAs of P-III metalloproteinase precursors, designated as stejnihagin-A and stejnihagin-B, respectively, from...
A novel C-type lectin-like protein, dabocetin, was purified from Daboia russellii siamensis venom. On SDS-polyacrylamide gel electrophoresis, it showed a single band with an apparent molecular weight of 28kDa and two distinct bands with the apparent molecular weights of 15.0kDa and 14.5kDa under non-reducing and reducing conditions, respectively. cDNA clones containing the coding sequences for dabocetin...
A novel protein, named BAS-AH, was purified and characterized from the skin of the toad Bufo andrewsi. BAS-AH is a single chain protein and the apparent molecular weight is about 63kDa as judged by SDS-PAGE. BAS-AH was determined to bind heme (0.89mol heme/mol protein) as determined by pyridine haemochrome analysis. Fifty percentage cytotoxic concentration (CC 50 ) of BAS-AH on C8166 cells...
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