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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
Strategies for the structural analysis of multi-protein complexes: Lessons from the 3D-Repertoire project.
J. Struct. Biol. 175, 147-158 (2011)
Structural studies of multi-protein complexes, whether by X-ray diffraction, scattering, NMR spectroscopy or electron microscopy, require stringent quality control of the component samples. The inability to produce 'keystone' subunits in a soluble and correctly folded form is a serious impediment to the reconstitution of the complexes. Co-expression of the components offers a valuable alternative to the expression of single proteins as a route to obtain sufficient amounts of the sample of interest. Even in cases where milligram-scale quantities of purified complex of interest become available, there is still no guarantee that good quality crystals can be obtained. At this step, protein engineering of one or more components of the complex is frequently required to improve solubility, yield or the ability to crystallize the sample. Subsequent characterization of these constructs may be performed by solution techniques such as Small Angle X-ray Scattering and Nuclear Magnetic Resonance to identify 'well behaved' complexes. Herein, we recount our experiences gained at protein production and complex assembly during the European 3D Repertoire project (3DR). The goal of this consortium was to obtain structural information on multi-protein complexes from yeast by combining crystallography, electron microscopy, NMR and in silico modeling methods. We present here representative set case studies of complexes that were produced and analyzed within the 3DR project. Our experience provides useful insight into strategies that are more generally applicable for structural analysis of protein complexes.
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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
Protein complex validation for structural biology; Limited proteolysis; Crystallography; NMR; SAXS; KEOPS/EKC; MAK; RES complex; Dom34; Hbs1; Trm8; Trm81; Trm112
ISSN (print) / ISBN
1047-8477
e-ISSN
1047-8477
Zeitschrift
Journal of Structural Biology
Quellenangaben
Band: 175,
Heft: 2,
Seiten: 147-158
Verlag
Elsevier
Verlagsort
San Diego, CA
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed
Institut(e)
Institute of Structural Biology (STB)