Open AccessX-ray free-electron laser studies reveal correlated motion during isopenicillin N synthase catalysis
Author(s) -
Patrick Rabe,
Jos J. A. G. Kamps,
Kyle D. Sutherlin,
J. Linyard,
Pierre Aller,
Cindy C. Pham,
Hiroki Makita,
I.J. Clifton,
M.A. McDonough,
T.M. Leissing,
Denis Shutin,
Pauline A. Lang,
A. Butryn,
Jürgen Brem,
Sheraz Gul,
Franklin D. Fuller,
In-Sik Kim,
Mun Hon Cheah,
Thomas Fransson,
Asmit Bhowmick,
I.D. Young,
Lee J. O’Riordan,
Aaron S. Brewster,
Ilaria Pettinati,
Margaret Doyle,
Yasumasa Joti,
Shigeki Owada,
Kensuke Tono,
A. Batyuk,
Mark S. Hunter,
Roberto Alonso-Mori,
Uwe Bergmann,
R.L. Owen,
Nicholas K. Sauter,
Timothy D. W. Claridge,
Carol V. Robinson,
Vittal K. Yachandra,
Junko Yano,
Jan Kern,
Allen M. Orville,
Christopher J. Schofield
Publication year - 2021
Publication title -
science advances
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.928
H-Index - 146
ISSN - 2375-2548
DOI - 10.1126/sciadv.abh0250
Subject(s) - catalysis , chemistry , enzyme , atp synthase , active site , yield (engineering) , femtosecond , stereochemistry , photochemistry , laser , materials science , biochemistry , physics , optics , metallurgy
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.