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

Leary, JA; Schenauer, MR; Stefanescu, R; Andaya, A; Ruotolo, BT; Robinson, CV; Thalassinos, K; ... Hershey, JWB; + view all (2009) Methodology for Measuring Conformation of Solvent-Disrupted Protein Subunits using T-WAVE Ion Mobility MS: An Investigation into Eukaryotic Initiation Factors. J AM SOC MASS SPECTR , 20 (9) 1699 - 1706. 10.1016/j.jasms.2009.05.003.

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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. (J Am Soc Mass Spectrom 2009, 20, 1699-1706) (C) 2009 Published by Elsevier Inc. on behalf of American Society for Mass Spectrometry

Type: Article
Title: Methodology for Measuring Conformation of Solvent-Disrupted Protein Subunits using T-WAVE Ion Mobility MS: An Investigation into Eukaryotic Initiation Factors
DOI: 10.1016/j.jasms.2009.05.003
Keywords: TRANSLATION FACTOR EIF3, MASS-SPECTROMETRY, COMPLEXES, WATER, PEPTIDES, STATES
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology
URI: http://discovery.ucl.ac.uk/id/eprint/127862
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