Purification and Identification of a Synthesized Neuroexcitatory Peptide to Confirm its Refolded Structure Present in the Injected Venom of Conus catus Snails
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In this study, synthesized c4g was successfully purified using reverse phase high performance liquid chromatography (RV-HPLC). After c4g fractions containing the major peak were combined and concentrated, samples of it were taken to Cal Tech to identify the mass of the major peak. Once a mass of 3028.5 ?2 Da was observed in the purified peak, the c4g peptide was refolded using oxidized and reduced glutathione, EDTA and Tris-HCL. If a successfully refolded c4g peptide mass difference of approximately 6 Da is observed, then a molecular target will be closer to being confirmed. Native c4g taken from different C. catus snails IV is also fingerprinted and later purified using RV-HPLC. Intraspecific variation of injected venom is seen among the snails. This is an important observation to make in order to determine which venoms can be combined and which venoms must be kept separate to maximize the purity and concentration of native c4g. When enough native c4g is purified and confirmed through MALDI-ToF MS, its physiological effects can be compared to its synthetic counterpart using the zebrafish motility assay or using the patch clamping technique.