Snake venom proteins identification by nano high performance liquid chromatography tandem mass spectrometry

Unrestricted In this study, a modified on-column frit preparation method for fused silica capillaries was used to analyze a venom mixture which contained the milkings from approximately ten snakes of the Agkistrodon contortrix contortrix (southern copperhead) species. One hundred and twenty-three proteins were identified by in-gel digestion coupled with nano-HPLC MS. Among them, twenty proteins matched the references and databases identified as snake venom proteins. In addition, de novo sequencing is applied to analyze the collision-induced dissociation (CID) data generated by in-solution digestion coupled with nano-HPLC MS. Twenty-three proteins matched the databases and references previously identified as snake venom proteins.; Because of insufficient sequences in the database, many peptides and hypothetical proteins could not be matched to the venom proteins. Some venom proteins we identified were found in other species. To build up an intact knowledge base to help toxicant study and clinical treatment, this thesis provided a reference for further protein purification, characterization, and function studies