Abstract
BackgroundSnake venoms are complex mixtures of toxic proteins or peptides encoded by various gene families that function synergistically to incapacitate prey. In the present study, in order to unravel the proteomic repertoire of Deinagkistrodon acutus venom, some trace abundance components were analyzed.MethodsShotgun proteomic approach combined with shotgun nano-LC-ESI-MS/MS were employed to characterize the medically important D. acutus venom, after collected samples were enriched with the combinatorial peptide ligand library (CPLL).ResultsThis avenue helped us find some trace components, undetected before, in D. acutus venom. The results indicated that D. acutus venom comprised 84 distinct proteins from 10 toxin families and 12 other proteins. These results are more than twice the number of venom components obtained from previous studies, which were only 29 distinct proteins obtained through RP-HPLC for the venom of the same species. The present results indicated that in D. acutus venom, the most abundant components (66.9%) included metalloproteinases, serine proteinases, and C-type lectin proteins; the medium abundant components (13%) comprised phospholipases A2 (PLA2) and 5’-nucleotidases and nucleases; whereas least abundant components (6%) were aminopeptidases, L-amino acid oxidases (LAAO), neurotoxins and disintegrins; and the trace components. The last were undetected before the use of conventional shotgun proteomics combined with shotgun nano-LC-ESI-MS/MS, such as cysteine-rich secretory proteins Da-CRPa, phospholipases B-like 1, phospholipases B (PLB), nerve growth factors (NGF), glutaminyl-peptide cyclortransferases (QC), and vascular non-inflammatory molecules 2 (VNN2).ConclusionThese findings demonstrated that the CPLL enrichment method worked well in finding the trace toxin proteins in D. acutus venom, in contrast with the previous venomic characterization of D. acutus by conventional LC-MS/MS. In conclusion, this approach combined with the CPLL enrichment was effective for allowing us to explore the hidden D. acutus venomic profile and extended the list of potential venom toxins.
Highlights
Snake venoms are complex mixtures of toxic proteins or peptides encoded by various gene families that function synergistically to incapacitate prey
We firstly explored an in-depth analysis of venomic of D. acutus by combinatorial peptide ligand library (CPLL) combined with shotgun nano-LCESI/MS/MS, which revealed a clear feature of protein distribution in D. acutus venom, which will facilitate to understand the mechanism of envenoming caused by D. acutus bites
The two most abundant proteins, such as snake venom metalloproteinase (SVMP) and serine protease (SVSP), were diminished in the elution fraction compared to the crude venom sample
Summary
Snake venoms are complex mixtures of toxic proteins or peptides encoded by various gene families that function synergistically to incapacitate prey. The five-paced viper, known as the Chinese moccasin (Deinagkistrodon acutus, once named as Agkistrodon acutus), is a unique member of the monotypic genus Deinagkistrodon of the Viperidae family, reaching up to 2 m in length and weighing over 5 kg. It is a highly venomous snake in China and its venom’s LD50 ranges from 0.04 mg/kg to 10.0 mg/kg SC [1]. Unraveling the biochemical composition of D. acutus venom is of great importance for understanding the molecular mechanism of envenoming led by its bites. Researches on a single component of snake venom do not completely reveal the clear composition of D. acutus venom
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