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Direct Identification of Tyrosine Sulfation by using Ultraviolet Photodissociation Mass Spectrometry

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

Sulfation is a common post-translational modification of tyrosine residues in eukaryotes; however, detection using traditional liquid chromatography-mass spectrometry (LC-MS) methods is challenging based on poor ionization efficiency in the positive ion mode and facile neutral loss upon collisional activation. In the present study, 193 nm ultraviolet photodissociation (UVPD) is applied to sulfopeptide anions to generate diagnostic sequence ions, which do not undergo appreciable neutral loss of sulfate even using higher energy photoirradiation parameters. At the same time, neutral loss of SO3 is observed from the precursor and charge-reduced precursor ions, a spectral feature that is useful for differentiating tyrosine sulfation from the nominally isobaric tyrosine phosphorylation. LC-MS detection limits for UVPD analysis in the negative mode were determined to be around 100 fmol for three sulfated peptides, caerulein, cionin, and leu-enkephalin. The LC-UVPD-MS method was applied for analysis of bovine fibrinogen, and its key sulfated peptide was confidently identified.

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Acknowledgments

The authors acknowledge support for this work by National Institute of Health grants R21GM099028 (to J.S.B.) and RO1HD056022 (to K.L.M.), the Robert A. Welch Foundation (F1155 to J.S.B.), and institutional funds from the Oklahoma Medical Research Foundation (to K.L.M).

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Authors and Affiliations

  1. Department of Chemistry, The University of Texas at Austin, 1 University Station A5300, Austin, TX, 78712, USA

    Michelle R. Robinson & Jennifer S. Brodbelt

  2. Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, 73104, USA

    Kevin L. Moore

  3. Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA

    Kevin L. Moore

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  1. Michelle R. Robinson
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Correspondence to Jennifer S. Brodbelt.

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Robinson, M.R., Moore, K.L. & Brodbelt, J.S. Direct Identification of Tyrosine Sulfation by using Ultraviolet Photodissociation Mass Spectrometry. J. Am. Soc. Mass Spectrom. 25, 1461–1471 (2014). https://doi.org/10.1007/s13361-014-0910-3

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