X-ray free-electron laser studies reveal correlated motion during isopenicillin N synthase catalysis

DOI: 10.1126/sciadv.abh0250

Authors: Patrick Rabe (University of Oxford) , Jos J. A. G. Kamps (University of Oxford; Diamond Light Source; Research Complex at Harwell) , Kyle D. Sutherlin (Lawrence Berkeley National Laboratory) , James D. S. Linyard (University of Oxford) , Pierre Aller (Diamond Light Source; Research Complex at Harwell) , Cindy C. Pham (Lawrence Berkeley National Laboratory) , Mikako Makita (Lawrence Berkeley National Laboratory) , Ian Clifton (University of Oxford) , Michael A. Mcdonough (University of Oxford) , Thomas M. Leissing (University of Oxford) , Denis Shutin (University of Oxford) , Pauline A. Lang (University of Oxford) , Agata Butryn (Diamond Light Source; Research Complex at Harwell) , Jurgen Brem (University of Oxford) , Sheraz Gul (Lawrence Berkeley National Laboratory) , Franklin D. Fuller (Linac Coherent Light Source, SLAC National Accelerator Laboratory) , In-Sik Kim (Lawrence Berkeley National Laboratory) , Mun Hon Cheah (Uppsala University) , Thomas Fransson (University of Heidelberg) , Asmit Bhowmick (Lawrence Berkeley National Laboratory) , Iris D. Young (Lawrence Berkeley National Laboratory; University of California, San Francisco) , Lee O'Riordan (Lawrence Berkeley National Laboratory) , Aaron S. Brewster (Lawrence Berkeley National Laboratory) , Ilaria Pettinati (University of Oxford) , Margaret Doyle (Lawrence Berkeley National Laboratory) , Yasumasa Joti (Japan Synchrotron Radiation Research Institute; RIKEN SPring-8 Center) , Shigeki Owada (Japan Synchrotron Radiation Research Institute; RIKEN SPring-8 Center) , Kensuke Tono (Japan Synchrotron Radiation Research Institute; RIKEN SPring-8 Center) , Alexander Batyuk (Linac Coherent Light Source, SLAC National Accelerator Laboratory) , Mark S. Hunter (Linac Coherent Light Source, SLAC National Accelerator Laboratory) , Roberto Alonso-Mori (Linac Coherent Light Source, SLAC National Accelerator Laboratory) , Uwe Bergmann (Stanford PULSE Institute, SLAC National Accelerator Laboratory; University of Wisconsin–Madison) , Robin L. Owen (Diamond Light Source) , Nicholas K. Sauter (Lawrence Berkeley National Laboratory) , Timothy D. W. Claridge (University of Oxford) , Carol V. Robinson (University of Oxford) , Vittal K. Yachandra (Lawrence Berkeley National Laboratory) , Junko Yano (Lawrence Berkeley National Laboratory) , Jan F. Kern (Lawrence Berkeley National Laboratory) , Allen M. Orville (Diamond Light Source; Research Complex at Harwell) , Christopher J. Schofield (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Science Advances , VOL 7

State: Published (Approved)
Published: August 2021
Diamond Proposal Number(s): 23459 , 19458

Abstract: Isopenicillin N synthase (IPNS) catalyzes the unique reaction of L-δ-(α-aminoadipoyl)-L-cysteinyl-D-valine (ACV) with dioxygen giving isopenicillin N (IPN), the precursor of all natural penicillins and cephalosporins. X-ray free-electron laser studies including time-resolved crystallography and emission spectroscopy reveal how reaction of IPNS:Fe(II):ACV with dioxygen to yield an Fe(III) superoxide causes differences in active site volume and unexpected conformational changes that propagate to structurally remote regions. Combined with solution studies, the results reveal the importance of protein dynamics in regulating intermediate conformations during conversion of ACV to IPN. The results have implications for catalysis by multiple IPNS-related oxygenases, including those involved in the human hypoxic response, and highlight the power of serial femtosecond crystallography to provide insight into long-range enzyme dynamics during reactions presently impossible for nonprotein catalysts.

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength) , I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

Added On: 23/08/2021 10:54

Documents:
eabh0250.full.pdf

Discipline Tags:

Health & Wellbeing Biochemistry Catalysis Chemistry Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)