期刊
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
卷 24, 期 20, 页码 3033-3042出版社
WILEY
DOI: 10.1002/rcm.4732
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资金
- White Rose DTC EPSRC
- BBSRC [BB/E008070/1, BB/E012558/1]
- The Wellcome Trust
- The Leverhulme Trust
- Biotechnology and Biological Sciences Research Council [BB/E008070/1, BB/E012558/1] Funding Source: researchfish
We have combined ion mobility spectrometry mass spectrometry with tandem mass spectrometry to characterise large, non-covalently bound macromolecular complexes in terms of mass, shape (cross-sectional area) and stability (dissociation) in a single experiment. The results indicate that the quaternary architecture of a complex influences its residual shape following removal of a single subunit by collision-induced dissociation tandem mass spectrometry. Complexes whose subunits are bound to several neighbouring subunits to create a ring-like three-dimensional (3D) architecture undergo significant collapse upon dissociation. In contrast, subunits which have only a single neighbouring subunit within a complex retain much of their original shape upon complex dissociation. Specifically, we have determined the architecture of two transient, on-pathway intermediates observed during in vitro viral capsid assembly. Knowledge of the mass, stoichiometry and cross-sectional area of each viral assembly intermediate allowed us to model a range of potential structures based on the known X-ray structure of the coat protein building blocks. Comparing the cross-sectional areas of these potential architectures before and after dissociation provided tangible evidence for the assignment of the topologies of the complexes, which have been found to encompass both the 3-fold and the 5-fold symmetry axes of the final icosahedral viral shell. Such insights provide unique information about virus assembly pathways that could allow the design of anti-viral therapeutics directed at the assembly step. This methodology can be readily applied to the structural characterisation of many other non-covalently bound macromolecular complexes and their assembly pathways. Copyright (C) 2010 John Wiley & Sons, Ltd.
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