Constructing comprehensive venom proteome reference maps for integrative venomics

Abstract

Our results underscore that if sufficient pre-MS and MS efforts are applied, comprehensive venom maps can be achieved. And - equally important - dissociating the venom decomplexing steps from the protein identification process represents the key to achieving a quantitative and locus-resolved insight of the venom proteome.

What killed Karl Patterson Schmidt? Combined venom gland transcriptomic, venomic and antivenomic analysis of the South African green tree snake (the boomslang), Dispholidus typus

“…3, Supplementary Table S1) and peptide-centric tandem mass spectrometry-based bottom-up venomics [37], [38]. Nano-electrospray ionisation mass spectrometry (nESI-MS/MS) identified proteins belonging to 7 different snake venom protein families (Fig.…”

“…0.75 mg of crude, lyophilised venom was dissolved in 200 μL of 5% acetonitrile in MilliQ® (Millipore Co.) water containing 0.1% trifluoroacetic acid (TFA), centrifuged to remove debris, and separated by reverse-phase (RP) HPLC using a Teknokroma Europa Protein 300 C18 (0.4 cm × 25 cm, 5 μm particle size, 300 Å pore size) column and an LC 1100 High Pressure Gradient System (Agilent Technologies, Santa Clara, CA, USA) equipped with DAD detector and micro-Auto-sampler [37]. The flow rate was set to 1 mL/min and the column was developed with a linear gradient of 0.1% TFA in water (solution A) and acetonitrile (solution B) using the following column elution conditions: isocratically (5% B) for 5 min, followed by 5%–25% B for 10 min, 25%–45% B for 60 min, and 45%–70% for 10 min.…”

Functional characterizations of venom phenotypes in the eastern diamondback rattlesnake (Crotalus adamanteus) and evidence for expression-driven divergence in toxic activities among populations

“…The specific toxic molecules responsible for this age-specific effect, however, was unclear because a number of toxin families can affect the coagulation cascade (e.g., C-type lectins and phospholipases A 2 ). C-type lectins can possess significant anticoagulant activity, but these toxins were much more abundant in adults (Margres et al, 2015b), although variation in C-type lectin expression may not be apparent when using RP-HPLC to quantify protein abundances (Eichberg et al, 2015). C-type lectins and snake venom type II metalloproteinases have a similar molecular weight, making it difficult to differentiate these toxins in our SDS-PAGE analysis (Fig.…”

Strategies in ‘snake venomics’ aiming at an integrative view of compositional, functional, and immunological characteristics of venoms

“…Among advantages, a full pattern of sample decomplexation can be obtained in a single two-dimensional gel electrophoresis (2DE), from which information on the isoelectric point (pI, first dimension) and apparent molecular weight (Mw, second dimension) of the proteins can be readily determined for each spot. Moreover, the macromolecular organization of venom proteins can also be assessed by comparing 2DE separations run under non-reducing conditions in both directions versus non-reducing (first dimension)/reducing (second dimension) [31]. Also, it is possible to stain the gel not only for proteins, but also for conjugated moieties such as glycosylations or other post-translational modifications (PTMs) of interest [32, 33].…”