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Ultralow-resolution ab initio phasing of filamentous proteins: crystals from a six-Ig fragment of titin as a case study.
IGBMC, CNRS-INSERM-ULP, Illkirch, France; Department of Physics, Faculty of Science and Technologies, Nancy-University, Vandoeuvre-les-Nancy, France.
Division of Structural Biology, Biozentrum, University of Basel, Basel, Switzerland.
Division of Structural Biology, Biozentrum, University of Basel, Basel, Switzerland; School of Biological Sciences, Biosciences Building, University of Liverpool, Liverpool, England.
2008 (English)In: Acta Crystallographica Section D: Biological Crystallography, ISSN 0907-4449, E-ISSN 1399-0047, Vol. 64, no Pt 5, p. 478-486Article in journal (Refereed) Published
Abstract [en]

Low-resolution diffraction data (resolution below 12 angstroms) from crystals of a filamentous six-Ig fragment of titin, I65-I70, were used in ab initio phasing with the aim of calculating its lattice packing and molecular envelope. Filamentous molecules, characterized by marked anisometry and idiosyncratic crystal lattices, have not been addressed before using this methodology. In this study, low-resolution phasing (19-122 angstroms) successfully identified the region of the unit cell occupied by the molecule. Phase extension to a higher resolution (12 angstroms) yielded regions of high density that corresponded either to the positions of individual Ig domains or to zones of dense intermolecular contacts, hindering the identification of individual domains and the interpretation of electron-density maps in terms of a molecular model. This problem resulted from the acutely uneven packing of the molecules in the crystal and it was further accentuated by the presence of partially disordered regions in the molecule. Addition of low-resolution reflections with phases computed ab initio to those obtained experimentally using MIRAS improved the initial electron-density maps of the atomic model, demonstrating the generic utility of low-resolution phases for the structure-elucidation process, even when individual molecules cannot be resolved in the lattice.

Place, publisher, year, edition, pages
Wiley-Blackwell Publishing Inc., 2008. Vol. 64, no Pt 5, p. 478-486
Keywords [en]
ultra-low resolution ab initio phasing, filamentous proteins, titin, crystal packing
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:liu:diva-143669DOI: 10.1107/S0907444908003442ISI: 000255077900002PubMedID: 18453683Scopus ID: 2-s2.0-43249092361OAI: oai:DiVA.org:liu-143669DiVA, id: diva2:1165130
Available from: 2017-12-12 Created: 2017-12-12 Last updated: 2017-12-21Bibliographically approved

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von Castelmur, Eleonore

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