Identification of tyrosine sulfation in Conus pennaceus conotoxins α-PnIA and α-PnIB: Further investigation of labile sulfo- and phosphopeptides by electrospray, matrix-assisted laser desorption/ionization (MALDI) and atmospheric pressure MALDI mass spectrometry

Jean Luc Wolfender, Feixia Chu, Haydn Ball, Florence Wolfender, Michael Fainzilber, Michael A. Baldwin, Alma L. Burlingame

Research output: Contribution to journalArticlepeer-review

88 Scopus citations

Abstract

Liquid chromatography/electrospray ionization mass spectrometry was used to investigate the peptide composition of the venom of Conus pennaceus, a molluscivorous cone shell from the Red Sea. Based on observed M(r)S, this venom contained all known conotoxins previously isolated and identified from this species. Interestingly, the doubly protonated species of only two of these conotoxins, α-PnIA and α-PnIB, showed additional related ions at +40 m/z (+80 Da), indicating the presence of either sulfation or phosphorylation in both components. High-performance liquid chromatographic (HPLC) fractions containing these two conotoxins were examined by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry in both positive and negative ion modes, as well as by MALDI high-energy collision-induced dissociation. These experiments established the presence of a single sulfated tyrosine residue within both α-PnIA and α-PnIB. Hence their post-translationally modified sequences are GCCSLPPCAANNPDY(S)C-NH2 (α-PnIA) and GCCSLPPCALSNPDY(S)C-NH2 (α-PnIB). This assignment was supported by comparison of their mass spectral behavior with that of known sulfated and phosphorylated peptides. This data clarified further the distinguishing features of the ionization and fragmentation of such modified peptides. Selective disulfide folding of synthetic α-PnIB demonstrated that both sulfated and non-sulfated toxins co-elute on reversed-phase HPLC and that α-PnIB possesses the same disulfide connectivity as other 'classical' α-conotoxins reported previously.

Original languageEnglish (US)
Pages (from-to)447-454
Number of pages8
JournalJournal of Mass Spectrometry
Volume34
Issue number4
DOIs
StatePublished - 1999

Keywords

  • Atmospheric pressure matrix-assisted laser desorption/ionization
  • Conotoxin
  • Electrospray ionization
  • Matrix-assisted laser desorption/ionization
  • Sulfotyrosine

ASJC Scopus subject areas

  • Spectroscopy

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