Modelling multi-protein complexes using PELDOR distance measurements for rigid body minimisation experiments using XPLOR-NIH

Crystallographic and NMR approaches have provided a wealth of structural information about protein domains. However, often these domains are found as components of larger multi domain polypeptides or complexes. Orienting domains within such contexts can provide powerful new insight into their functi...

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Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 2014-12, Vol.70 (2-3), p.139-153
Main Authors: Hammond, Colin M., Owen-Hughes, Tom, Norman, David G.
Format: Article
Language:English
Subjects:
Chromatin
Computational Biology - methods
DEER
Electron Spin Resonance Spectroscopy - methods
Histone Chaperones - chemistry
Models, Molecular
MTSSLwizard
PELDOR
Software
Spin Labels
Vps75
XPLOR-NIH
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Summary:Crystallographic and NMR approaches have provided a wealth of structural information about protein domains. However, often these domains are found as components of larger multi domain polypeptides or complexes. Orienting domains within such contexts can provide powerful new insight into their function. The combination of site specific spin labelling and Pulsed Electron Double Resonance (PELDOR) provide a means of obtaining structural measurements that can be used to generate models describing how such domains are oriented. Here we describe a pipeline for modelling the location of thio-reactive nitroxyl spin locations to engineered sties on the histone chaperone Vps75. We then use a combination of experimentally determined measurements and symmetry constraints to model the orientation in which homodimers of Vps75 associate to form homotetramers using the XPLOR-NIH platform. This provides a working example of how PELDOR measurements can be used to generate a structural model.
ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2014.10.028