Screening for post-translational modifications in conotoxins using liquid chromatography/mass spectrometry: an important component of conotoxin discovery.
Mass spectrometry has emerged as an important technique for conotoxin analysis due to its capacity for selective, sensitive, information-rich analyses. Using liquid chromatography/mass spectrometry, Conus venom can be fractionated and the peptides surveyed for specific post-translational modifications, indicating those toxin components likely to have an important biological function. With Conus striatus and Conus victoriae venom as models, bromination, carboxylation and glycosylation modifications are identified through characteristics such as isotopic distribution and labile losses observed during mass spectrometric analysis. This modification screening approach enables the identification of a C. victoriae bromo-carboxy-conotoxin, designated vc5c, as a candidate for detailed mass spectrometric analysis. Using a cDNA sequence coupled with liquid chromatography/mass spectrometry and nanoelectrospray ionization-ion trap-mass spectrometry, the sequence of vc5c is determined to be ICCYPNXWCCD, where W is 6-bromotryptophan, X is gamma-carboxy glutamate and C is disulfide-linked cysteine. This represents the ninth T-superfamily (-CC-CC- scaffold) toxin that has been isolated from venom and characterized.
Jakubowski
Jennifer A
JA
Department of Chemistry and the Beckman Institute, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue 63-5, Urbana, IL 61801, USA.
Kelley
Wayne P
WP
Sweedler
Jonathan V
JV
D000595:Amino Acid Sequence; D000818:Animals; D001315:Australia; D002853:Chromatography, Liquid / Q000379:methods*; D020916:Conotoxins / Q000032:analysis / Q000235:genetics*; D052078:Conus Snail / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000032:analysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008978:Mollusk Venoms / Q000032:analysis*; D011499:Protein Processing, Post-Translational; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D020916:Conotoxins; D018076:DNA, Complementary; D008978:Mollusk Venoms