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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
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Leary, Julie A., Schenauer, Matthew R., Stefanescu, Raluca, Andaya, Armann, Ruotolo, Brandon T., Robinson, Carol V., Thalassinos, Konstantinos, Scrivens, James H., Sokabe, Masaaki and Hershey, John W. B. (2009) Methodology for measuring conformation of solvent-disrupted protein subunits using T-WAVE ion mobility MS: an investigation into eukaryotic initiation factors. Journal of The American Society for Mass Spectrometry, Vol.20 (No.9). pp. 1699-1706. doi:10.1016/j.jasms.2009.05.003
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Official URL: http://dx.doi.org/10.1016/j.jasms.2009.05.003
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
| Item Type: | Journal Article | ||||
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| Subjects: | Q Science > QD Chemistry Q Science > QC Physics |
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| Divisions: | Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010) | ||||
| Journal or Publication Title: | Journal of The American Society for Mass Spectrometry | ||||
| Publisher: | Springer New York LLC | ||||
| ISSN: | 1044-0305 | ||||
| Official Date: | September 2009 | ||||
| Dates: |
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| Volume: | Vol.20 | ||||
| Number: | No.9 | ||||
| Number of Pages: | 8 | ||||
| Page Range: | pp. 1699-1706 | ||||
| DOI: | 10.1016/j.jasms.2009.05.003 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Restricted or Subscription Access | ||||
| Funder: | NIH PHS, Human Frontier Science Program Fellowship | ||||
| Grant number: | GM073732, LT00575/2007 |
Data sourced from Thomson Reuters' Web of Knowledge
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