title: Fingerprinting redox and ligand states in haemprotein crystal structures using resonance Raman spectroscopy.
abstract: It is crucial to assign the correct redox and ligand states to crystal structures of proteins with an active redox centre to gain valid functional information and prevent the misinterpretation of structures. Single-crystal spectroscopies, particularly when applied in situ at macromolecular crystallography beamlines, allow spectroscopic investigations of redox and ligand states and the identification of reaction intermediates in protein crystals during the collection of structural data. Single-crystal resonance Raman spectroscopy was carried out in combination with macromolecular crystallography on Swiss Light Source beamline X10SA using cytochrome c' from Alcaligenes xylosoxidans. This allowed the fingerprinting and validation of different redox and ligand states, identification of vibrational modes and identification of intermediates together with monitoring of radiation-induced changes. This combined approach provides a powerful tool to obtain complementary data and correctly assign the true oxidation and ligand state(s) in redox-protein crystals.
authors.affiliation: School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, England.
authors.forename: Demet
authors.initials: D
authors.lastname: Kekilli
authors.affiliation: Swiss Light Source, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Florian S N
authors.initials: FS
authors.lastname: Dworkowski
authors.affiliation: Swiss Light Source, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Pompidor
authors.affiliation: Swiss Light Source, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Martin R
authors.initials: MR
authors.lastname: Fuchs
authors.affiliation: Department of Chemistry and Biochemistry, Eastern Oregon University, La Grande, OR 97850-2899, USA.
authors.forename: Colin R
authors.initials: CR
authors.lastname: Andrew
authors.affiliation: Institute of Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, England.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Antonyuk
authors.affiliation: Institute of Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, England.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Strange
authors.affiliation: Institute of Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, England.
authors.forename: Robert R
authors.initials: RR
authors.lastname: Eady
authors.affiliation: Institute of Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, England.
authors.forename: S Samar
authors.initials: SS
authors.lastname: Hasnain
authors.affiliation: School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, England.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Hough
mesh_terms: D000421:Alcaligenes / Q000737:chemistry; D018360:Crystallography, X-Ray / Q000379:methods*; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism; D006420:Hemeproteins / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D013059:Spectrum Analysis, Raman
chemical_list: D006420:Hemeproteins; D008024:Ligands; D045304:Cytochromes c
keywords: cytochrome c′; haemproteins; ligand states; redox states; resonance Raman spectroscopy