Streamlining the Analysis of Proteins from Snake Venom

Area of Science:

• Biochemistry
• Proteomics
• Toxicology

Background:

• Snake venom contains complex protein mixtures with therapeutic potential.
• Characterizing individual venom components presents significant technical challenges.
• Advanced analytical techniques are needed to overcome these limitations.

Purpose of the Study:

• To develop and demonstrate novel mass spectrometry workflows for analyzing complex snake venom.
• To characterize individual proteins and their modifications within King cobra venom.
• To showcase the utility of native and top-down mass spectrometry for venom proteomics.

Main Methods:

• Coupling ion mobility spectrometry with electron capture dissociation for protein separation and charge reduction.
• Performing top-down glycomic analysis on a specific King cobra venom toxin.
• Utilizing temperature-controlled nanoelectrospray mass spectrometry for top-down sequence analysis of disulfide-rich toxins.

Main Results:

• Successfully resolved complex mass spectra of multiple venom proteins (55-127 kDa) using ion mobility and electron capture dissociation.
• Identified a fucosylated complex glycan on a 25.5 kDa King cobra venom toxin.
• Determined the sequence of a β-cardiotoxin resistant to conventional fragmentation methods.

Conclusions:

• Native and top-down mass spectrometry workflows are effective for analyzing complex snake venom proteomes.
• These advanced MS methods enable detailed characterization of venom proteins and their post-translational modifications.
• The presented methodologies offer promising solutions for snake venom research and drug discovery.