期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 306, 期 3, 页码 389-396出版社
ACADEMIC PRESS LTD
DOI: 10.1006/jmbi.2000.4383
关键词
hydrogen/deuterium exchange; virus; capsid; protein folding; dynamics
资金
- NCI NIH HHS [P30 CA13148-27] Funding Source: Medline
- NCRR NIH HHS [S10RR11329] Funding Source: Medline
The dynamics of proteins within large cellular assemblies are important in the molecular transformations that are required for macromolecular synthesis, transport, and metabolism. The capsid expansion (maturation) accompanying DNA packaging in the dsDNA bacteriophage P22 represents an experimentally accessible case of such a transformation. A novel method, based on hydrogen-deuterium exchange was devised to investigate the dynamics of capsid expansion. Mass spectrometric detection of deuterium incorporation allows for a sensitive and quantitative determination of hydrogen-deuterium exchange dynamics irrespective of the size of the assembly. Partial digestion of the exchanged protein with pepsin allows for region-specific assignment of the exchange. Procapsids and mature capsids were probed under native and slightly denaturing conditions. These experiments revealed regions that exhibit different degrees of flexibility in the procapsid and in the mature capsid. In addition, exchange and deuterium trapping during the process of expansion itself was observed and allowed for the identification of segments of the protein subunit that become buried or stabilized as a result of expansion. This approach may help to identify residues participating in macromolecular transformations and uncover novel patterns and hierarchies of interactions that determine functional movements within molecular machines. (C) 2001 Academic Press.
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