Screening Method for the Discovery of Potential Bioactive Cysteine-Containing Peptides Using 3D Mass Mapping

Area of Science:

• Biochemistry
• Pharmacology
• Proteomics

Background:

• Animal venoms and toxins are rich sources of bioactive peptides.
• Disulfide bonds enhance peptide stability and pharmacological activity.
• Discovering novel disulfide-containing peptides is crucial for drug development.

Purpose of the Study:

• To develop and apply a novel workflow for identifying disulfide-containing peptides.
• To screen frog skin secretions for new bioactive peptides with pharmacologic relevance.
• To utilize 3D mass mapping for distinguishing sulfur-containing peptides.

Main Methods:

• Developed a 3D mass mapping workflow utilizing mass, normalized isotopic shift (NIS), and normalized mass defect (NMD).
• Investigated skin secretions from Odorrana schmackeri and Bombina variegata.
• Separated peptides using nanoflow LC, acquired high-resolution scans, and calculated NMD and NIS.
• Selected candidate peptides with low NMD and high NIS for de novo sequencing.

Main Results:

• Successfully differentiated sulfur-containing peptides from those lacking sulfur using 3D mass mapping.
• Identified several novel peptides containing inter- and intra-molecular disulfide bonds.
• Demonstrated the utility of the workflow in discovering new bioactive peptides from complex biological samples.

Conclusions:

• The 3D mass mapping workflow is effective for discovering novel disulfide-containing peptides.
• Frog skin secretions are a promising source of pharmacologically relevant peptides.
• This approach advances the discovery of peptide-based therapeutics.