title: Pushing the limits of sulfur SAD phasing: de novo structure solution of the N-terminal domain of the ectodomain of HCV E1.
abstract: Single-wavelength anomalous dispersion of S atoms (S-SAD) is an elegant phasing method to determine crystal structures that does not require heavy-atom incorporation or selenomethionine derivatization. Nevertheless, this technique has been limited by the paucity of the signal at the usual X-ray wavelengths, requiring very accurate measurement of the anomalous differences. Here, the data collection and structure solution of the N-terminal domain of the ectodomain of HCV E1 from crystals that diffracted very weakly is reported. By combining the data from 32 crystals, it was possible to solve the sulfur substructure and calculate initial maps at 7 Å resolution, and after density modication and phase extension using a higher resolution native data set to 3.5 Å resolution model building was achievable.
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
authors.forename: Kamel
authors.initials: K
authors.lastname: El Omari
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
authors.forename: Oleg
authors.initials: O
authors.lastname: Iourin
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
authors.forename: Jan
authors.initials: J
authors.lastname: Kadlec
authors.affiliation: Diamond Light Source Ltd, Diamond House, Harwell Science and Innovation Campus, Didcot OX11 0DE, England.
authors.forename: Richard
authors.initials: R
authors.lastname: Fearn
authors.affiliation: Diamond Light Source Ltd, Diamond House, Harwell Science and Innovation Campus, Didcot OX11 0DE, England.
authors.forename: David R
authors.initials: DR
authors.lastname: Hall
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
authors.forename: Karl
authors.initials: K
authors.lastname: Harlos
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Grimes
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
authors.forename: David I
authors.initials: DI
authors.lastname: Stuart
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D016174:Hepacivirus / Q000737:chemistry*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: C087035:E1 protein, Hepatitis C virus; D014759:Viral Envelope Proteins
keywords: HCV; envelope glycoprotein E1; sulfur SAD
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