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Structure and Mechanism of a Viral Collagen Prolyl Hydroxylase

DOI: 10.1021/acs.biochem.5b00789 DOI Help
PMID: 26368022 PMID Help

Authors: James E. Longbotham (University of Manchester) , Colin Levy (School of Chemistry & Manchester Interdisciplinary Biocentre, The University of Manchester) , Linus O. Johannissen (University of Manchester) , Hanna Tarhonskaya (University of Oxford) , Shuo Jiang (University of Oxford) , Christoph Loenarz (University of Oxford) , Emily Flashman (University of Oxford) , Sam Hay (University of Manchester) , Christopher J. Schofield (University of Oxford) , Nigel S. Scrutton (University of Manchester)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Biochemistry , VOL 54 (39) , PAGES 6093 - 6105

State: Published (Approved)
Published: October 2015
Diamond Proposal Number(s): 7146

Open Access Open Access

Abstract: The Fe(II)- and 2-oxoglutarate (2-OG)-dependent dioxygenases comprise a large and diverse enzyme superfamily the members of which have multiple physiological roles. Despite this diversity, these enzymes share a common chemical mechanism and a core structural fold, a double-stranded β-helix (DSBH), as well as conserved active site residues. The prolyl hydroxylases are members of this large superfamily. Prolyl hydroxylases are involved in collagen biosynthesis and oxygen sensing in mammalian cells. Structural− mechanistic studies with prolyl hydroxylases have broader implications for understanding mechanisms in the Fe(II)- and 2-OGdependent dioxygenase superfamily. Here, we describe crystal structures of an N-terminally truncated viral collagen prolyl hydroxylase (vCPH). The crystal structure shows that vCPH contains the conserved DSBH motif and iron binding active site residues of 2-OG oxygenases. Molecular dynamics simulations are used to delineate structural changes in vCPH upon binding its substrate. Kinetic investigations are used to report on reaction cycle intermediates and compare them to the closest homologues of vCPH. The study highlights the utility of vCPH as a model enzyme for broader mechanistic analysis of Fe(II)- and 2-OGdependent dioxygenases, including those of biomedical interest.

Subject Areas: Biology and Bio-materials

Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

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