Characterization of the two myotoxin a isomers from the prairie rattlesnake ( Crotalus viridis viridis) by capillary zone electrophoresis and fluorescence quenching studies

Abstract

The two myotoxin α isomers from the venom of the prairie rattlesnake Crotalus viridis viridis have different isoelectric points, as determined by capillary zone electrophoresis. The p I values are 10.50 and 10.57, respectively, and both are higher than the previously reported p I value for myotoxin α. The difference in the isoelectric points between the two isomers is attributed to altered surface charge as a result of the conformational change in myotoxin α. Both isomers exist in crude venom, discounting the possibility that they are artifacts formed during the purification process. Fluorescence quenching of myotoxin α reveals heterogeneity of the tryptophans, possibly due to different environments. The fraction of the total tryptophan fluorescence quenched by iodide is 81% and is attributed to solvent-accessible tryptophan residues at the protein surface. The 19% non-quenchable tryptophans probably represent residues that are shielded from the solvent exposure. The ratio of buried to exposed tryptophans is similar to the ratio of isomers seen by capillary zone electrophoresis and reverse-phase high-performance liquid chromatography ( C. 1:4).