X-ray crystallographic studies of substrate binding to aristolochene synthase suggest a metal ion binding sequence for catalysis

Shishova, Ekaterina Y., Yu, Fanglei, Miller, David James, Faraldos, Juan A., Zhao, Yuxin X., Coates, Robert M., Allemann, Rudolf Konrad ORCID: https://orcid.org/0000-0002-1323-8830, Cane, David E. and Christianson, David W. 2008. X-ray crystallographic studies of substrate binding to aristolochene synthase suggest a metal ion binding sequence for catalysis. Journal of Biological Chemistry 283 (22) , pp. 15431-15439. 10.1074/jbc.M800659200

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Abstract

The universal sesquiterpene precursor, farnesyl diphosphate (FPP), is cyclized in an Mg2+-dependent reaction catalyzed by the tetrameric aristolochene synthase from Aspergillus terreus to form the bicyclic hydrocarbon aristolochene and a pyrophosphate anion (PPi) coproduct. The 2.1-Å resolution crystal structure determined from crystals soaked with FPP reveals the binding of intact FPP to monomers A-C, and the binding of PPi and Mg2+B to monomer D. The 1.89-Å resolution structure of the complex with 2-fluorofarnesyl diphosphate (2F-FPP) reveals 2F-FPP binding to all subunits of the tetramer, with Mg2+Baccompanying the binding of this analogue only in monomer D. All monomers adopt open activesite conformations in these complexes, but slight structural changes in monomers C and D of each complex reflect the very initial stages of a conformational transition to the closed state. Finally, the 2.4-Å resolution structure of the complex with 12,13-difluorofarnesyl diphosphate (DF-FPP) reveals the binding of intact DF-FPP to monomers A-C in the open conformation and the binding of PPi, Mg2+B, and Mg2+C to monomer D in a predominantly closed conformation. Taken together, these structures provide 12 independent “snapshots” of substrate or product complexes that suggest a possible sequence for metal ion binding and conformational changes required for catalysis.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Chemistry Cardiff Catalysis Institute (CCI)
Publisher:	American Society for Biochemistry and Molecular Biology
ISSN:	0021-9258
Funders:	EPSRC
Last Modified:	17 Oct 2022 09:54
URI:	https://orca.cardiff.ac.uk/id/eprint/6121

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