5KWF

Joint X-ray Neutron Structure of Cholesterol Oxidase

Summary for 5KWF

• Entry DOI: 10.2210/pdb5kwf/pdb
• Descriptor: Cholesterol oxidase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• Functional Keywords: oxidoreductase, isomerase
• Biological source: Streptomyces sp. (strain SA-COO)
• Cellular location: Secreted: P12676
• Total number of polymer chains: 1
• Total formula weight: 56584.09

Authors

Golden, E.,Vrielink, A.,Meilleur, F.,Blakeley, M. (deposition date: 2016-07-18, release date: 2017-02-01, Last modification date: 2024-03-06)

Primary citation

Golden, E.,Yu, L.J.,Meilleur, F.,Blakeley, M.P.,Duff, A.P.,Karton, A.,Vrielink, A.
An extended N-H bond, driven by a conserved second-order interaction, orients the flavin N5 orbital in cholesterol oxidase.
Sci Rep, 7:40517-40517, 2017

PubMed Abstract: The protein microenvironment surrounding the flavin cofactor in flavoenzymes is key to the efficiency and diversity of reactions catalysed by this class of enzymes. X-ray diffraction structures of oxidoreductase flavoenzymes have revealed recurrent features which facilitate catalysis, such as a hydrogen bond between a main chain nitrogen atom and the flavin redox center (N5). A neutron diffraction study of cholesterol oxidase has revealed an unusual elongated main chain nitrogen to hydrogen bond distance positioning the hydrogen atom towards the flavin N5 reactive center. Investigation of the structural features which could cause such an unusual occurrence revealed a positively charged lysine side chain, conserved in other flavin mediated oxidoreductases, in a second shell away from the FAD cofactor acting to polarize the peptide bond through interaction with the carbonyl oxygen atom. Double-hybrid density functional theory calculations confirm that this electrostatic arrangement affects the N-H bond length in the region of the flavin reactive center. We propose a novel second-order partial-charge interaction network which enables the correct orientation of the hydride receiving orbital of N5. The implications of these observations for flavin mediated redox chemistry are discussed.

PubMed: 28098177
DOI: 10.1038/srep40517

Experimental method

NEUTRON DIFFRACTION (2.214 Å)
X-RAY DIFFRACTION (1.499 Å)