Methodology for Measuring Conformation of Solvent-Disrupted Protein Subunits using T-WAVE Ion Mobility MS: An Investigation into Eukaryotic Initiation Factors

Julie A. Leary, Matthew R. Schenauer, Raluca Stefanescu, Armann Andaya, Brandon T. Ruotolo, Carol V. Robinson, Konstantinos Thalassinos, James Scrivens, Masaaki Sokabe, John W.B. Hershey

    Research output: Contribution to journalArticle

    45 Citations (Scopus)

    Abstract

    The methodology developed in the research presented herein makes use of chaotropic solvents to gently dissociate subunits from an intact macromolecular complex and subsequently allows for the measurement of collision cross section (CCS) for both the recombinant (R-eIF3k) and solvent dissociated form of the subunit (S-eIF3k). In this particular case, the k subunit from the eukaryotic initiation factor 3 (eIF3) was investigated in detail. Experimental and theoretical CCS values show both the recombinant and solvent disrupted forms of the protein to be essentially the same. The ultimate goal of the project is to structurally characterize all the binding partners of eIF3, determine which subunits interact directly, and investigate how subunits may change conformation when they form complexes with other proteins. Research presented herein is the first report showing retention of solution conformation of a protein as evidenced by CCS measurements of both recombinant and solvent disrupted versions of the same protein.

    Original languageEnglish
    Pages (from-to)1699-1706
    Number of pages8
    JournalJournal of the American Society for Mass Spectrometry
    Volume20
    Issue number9
    DOIs
    Publication statusPublished - 1 Sep 2009

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