HK97 Maturation Studied by Crystallography and H/2H Exchange Reveals the Structural Basis for Exothermic Particle Transitions

JOURNAL OF MOLECULAR BIOLOGY

卷:397

Article

Gertsman, Ilya^1,2  Komives, Elizabeth A.²  Johnson, John E.^1,2 

[1] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

[2] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92037 USA

关键词: virus assembly;    X-ray crystallography;    hydrogen/deuterium exchange;    mass spectrometry;    protein folding;   

DOI  :  10.1016/j.jmb.2010.01.016

来源: Elsevier

【 摘 要 】

HK97 is an exceptionally amenable system for characterizing major conformational changes associated with capsid maturation in double-stranded DNA bacteriophage. HK97 undergoes a capsid expansion of similar to 20%, accompanied by major subunit rearrangements during genome packaging. A previous 3.44-angstrom-resolution crystal structure of the mature capsid Head II and cryo-electron microscopy studies of other intermediate expansion forms of HK97 suggested that, primarily, rigid-body movements facilitated the maturation process. We recently reported a 3.65-angstrom-resolution structure of the preexpanded particle form Prohead II (P-II) and found that the capsid subunits undergo significant refolding and twisting of the tertiary structure to accommodate expansion. The P-II study focused on major twisting motions in the P-domain and on refolding of the spine helix during the transition. Here we extend the crystallographic comparison between and Head II, characterizing the refolding events occurring in each of the four major domains of the capsid subunit and their effect on quaternary structure stabilization. In addition, hydrogen/deuterium exchange, coupled to mass spectrometry, was used to characterize the structural dynamics of three distinct capsid intermediates: P-II, Expansion Intermediate, and the nearly mature Head I. Differences in the solvent accessibilities of the seven quasi-equivalent capsid subunits, attributed to differences in secondary and quaternary structures, were observed in P-II. Nearly all differences in solvent accessibility among subunits disappear after the first transition to Expansion Intermediate. We show that most of the refolding is coupled to this transformation, an event associated with the transition from asymmetric to symmetric hexamers. (C) 2010 Elsevier Ltd. All rights reserved.

【 授权许可】

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