Journal of the American Society for Mass Spectrometry

, Volume 18, Issue 8, pp 1396–1404

Sequencing of T-superfamily conotoxins from Conus virgo: Pyroglutamic acid identification and disulfide arrangement by MALDI mass spectrometry

Authors

  • Amit Kumar Mandal
    • Molecular Biophysics UnitIndian Institute of Science
  • Mani Ramakrishnan Santhana Ramasamy
    • National Center for Biological Sciences
  • Varatharajan Sabareesh
    • Molecular Biophysics UnitIndian Institute of Science
  • Matthew E. Openshaw
    • Shimadzu Biotech
  • Kozhalmannom S. Krishnan
    • National Center for Biological Sciences
    • Department of Biological SciencesTata Institute of Fundamental Research
    • Molecular Biophysics UnitIndian Institute of Science
Articles

DOI: 10.1016/j.jasms.2007.04.009

Cite this article as:
Mandal, A.K., Ramasamy, M.R.S., Sabareesh, V. et al. J Am Soc Mass Spectrom (2007) 18: 1396. doi:10.1016/j.jasms.2007.04.009

Abstract

De novo mass spectrometric sequencing of two Conus peptides, Vi1359 and Vi1361, from the vermivorous cone snail Conus virgo, found off the southern Indian coast, is presented. The peptides, whose masses differ only by 2 Da, possess two disulfide bonds and an amidated C-terminus. Simple chemical modifications and enzymatic cleavage coupled with matrix assisted laser desorption ionization (MALDI) mass spectrometric analysis aided in establishing the sequences of Vi1359, ZCCITIPECCRI-NH2, and Vi1361, ZCCPTMPECCRI-NH2, which differ only at residues 4 and 6 (Z=pyroglutamic acid). The presence of the pyroglutamyl residue at the N-terminus was unambiguously identified by chemical hydrolysis of the cyclic amide, followed by esterification. The presence of Ile residues in both the peptides was confirmed from high-energy collision induced dissociation (CID) studies, using the observation of wn- and dn-ions as a diagnostic. Differential cysteine labeling, in conjunction with MALDI-MS/MS, permitted establishment of disulfide connectivity in both peptides as Cys2-Cys9 and Cys3-Cys10. The cysteine pattern clearly reveals that the peptides belong to the class of T-superfamily conotoxins, in particular the T-1 superfamily.

Download to read the full article text

Copyright information

© American Society for Mass Spectrometry 2007