Observation of Heterogeneous Gene Products by FT-ICR MS

D. H L Robertson, S. C C Wong, Robert J. Beynon, Jane L. Hurst, Simon J. Gaskell

Research output: Contribution to journalArticlepeer-review

Abstract

The ability of FT-ICR MS to resolve isotopic variants of intact proteins for each of the charge states formed by electrospray ionization offers a sensitive, rapid method for detecting "low mass" heterogeneity, where this is defined as the presence of structural variants differing in mass by 2 Da or less. Such heterogeneity may reflect biological or chemical modifications of structure or may result from the coexpression of related proteins from a multi-gene family. In the analytical approach described here, comparisons are made between observed isotopic distributions and those expected for predicted protein sequences. Close agreement is demonstrated for a homogeneous model protein, and the utility of the method has been evaluated in the study of mouse major urinary proteins (MUPs), a group of closely related sequences. Divergence of the experimental isotopic distribution from distributions predicted for known MUP sequences can be explained, in quantitative terms, by the coexpression of closely related sequences. This approach provides a facile method for the assessment of protein homogeneity and for the detection of structural variants, without recourse to proteolytic digestion and analysis of the resulting products. © 2008 American Society for Mass Spectrometry.
Original languageEnglish
Pages (from-to)103-110
Number of pages7
JournalJournal of the American Society for Mass Spectrometry
Volume19
Issue number1
DOIs
Publication statusPublished - Jan 2008

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