Conformational flexibility of the C terminus with implications for substrate binding and catalysis revealed in a new crystal form of deacetoxycephalosporin C synthase

Journal of Molecular Biology
Linda M OsterI Andersson

Abstract

Deacetoxycephalosporin C synthase (DAOCS) from Streptomyces clavuligerus catalyses the oxidative ring expansion of the penicillin nucleus into the nucleus of cephalosporins. The reaction requires dioxygen and 2-oxoglutarate as co-substrates to create a reactive iron-oxygen intermediate from a ferrous iron in the active site. The active enzyme is monomeric in solution. The structure of DAOCS was determined earlier from merohedrally twinned crystals where the last four C-terminal residues (308-311) of one molecule penetrate the active site of a neighbouring molecule, creating a cyclic trimeric structure in the crystal. Shortening the polypeptide chain from the C terminus by more than four residues diminishes activity. Here, we describe a new crystal form of DAOCS in which trimer formation is broken and the C-terminal arm is free. These crystals show no signs of twinning, and were obtained from DAOCS labelled with an N-terminal His-tag. The modified DAOCS is catalytically active. The free C-terminal arm protrudes into the solvent, and the C-terminal domain (residues 268-299) is rotated by about 16 degrees towards the active site. The last 12 residues (300-311) are disordered. Structures for various enzyme-substrate and enzyme-prod...Continue Reading

References

Jan 22, 1998·Current Opinion in Structural Biology·C J SchofieldP Roach
Sep 2, 1998·Nature·K ValegårdI Andersson
Oct 3, 1998·Acta Crystallographica. Section D, Biological Crystallography·A T BrüngerG L Warren
Mar 25, 1999·Acta Crystallographica. Section D, Biological Crystallography·R M Esnouf
Mar 25, 1999·Acta Crystallographica. Section D, Biological Crystallography·C R KissingerD B Fogel
May 1, 1999·Journal of Molecular Biology·M D LloydR Bhikhabhai
Jan 7, 2000·Biochemical and Biophysical Research Communications·H J LeeC J Schofield
Mar 30, 2001·The Journal of Biological Chemistry·H J LeeC J Schofield
May 16, 2001·Journal of Molecular Biology·H J LeeC J Schofield
Jul 5, 2001·Cellular and Molecular Life Sciences : CMLS·A DubusJ M Frere
Jul 27, 2001·Acta Crystallographica. Section D, Biological Crystallography·M Harris, T A Jones
Nov 22, 2001·Acta Crystallographica. Section D, Biological Crystallography·A C Terwisscha van ScheltingaI Andersson
Dec 12, 2001·European Journal of Biochemistry·I J CliftonC J Schofield
Feb 14, 2002·Structure·Rupert C WilmouthChristopher J Schofield
Nov 7, 1996·Chemical Reviews·Lawrence Que, Raymond Y. N. Ho
Mar 14, 2002·Biochemical and Biophysical Research Communications·Hwei-Jen LeeMatthew D Lloyd
Jun 1, 2002·Current Opinion in Chemical Biology·Matthew J Ryle, Robert P Hausinger
Jun 6, 2002·European Journal of Biochemistry·Sarah J LipscombMatthew D Lloyd
Nov 15, 2002·Proceedings of the National Academy of Sciences of the United States of America·Charles E DannJohann Deisenhofer
Nov 26, 2002·The Journal of Biological Chemistry·Jonathan M ElkinsChristopher J Schofield
Oct 24, 2003·Acta Crystallographica. Section D, Biological Crystallography·Anke C Terwisscha van ScheltingaInger Andersson
Jan 14, 2004·Nature Structural & Molecular Biology·Karin ValegårdInger Andersson
Jan 22, 2004·The Journal of Biological Chemistry·Matthew D LloydChristopher J Schofield
Sep 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·UNKNOWN Collaborative Computational Project, Number 4
May 1, 1997·Acta Crystallographica. Section D, Biological Crystallography·G N MurshudovE J Dodson
Jan 1, 1997·Methods in Enzymology·E A Merritt, D J Bacon
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor
Jan 1, 1997·Methods in Enzymology·Todd O Yeates

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Citations

May 20, 2005·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·K S HewitsonC J Schofield
Apr 14, 2007·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Vincent Purpero, Graham R Moran
Nov 13, 2012·Current Opinion in Structural Biology·WeiShen AikChristopher J Schofield
Jul 25, 2009·Current Opinion in Chemical Biology·Panqing He, Graham R Moran
Apr 19, 2017·Chemical Reviews·Kyle L DunbarChristian Hertweck
Mar 4, 2006·Journal of Inorganic Biochemistry·Ian J CliftonChristopher J Schofield
Jul 26, 2014·Natural Product Reports·Russell Cox
May 29, 2018·Natural Product Reports·Patrick RabeChristopher T Lohans
Apr 17, 2016·Applied and Environmental Microbiology·Nataraj BalakrishnanMicheal Durairaaj
Nov 9, 2012·Natural Product Reports·Refaat B HamedChristopher J Schofield

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