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An active site–tail interaction in the structure of hexahistidine‐tagged Thermoplasma acidophilum citrate synthase
Author(s) -
Murphy Jesse R.,
Donini Stefano,
Kappock T. Joseph
Publication year - 2015
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x15015939
Subject(s) - active site , thermoplasma acidophilum , citrate synthase , chemistry , binding site , stereochemistry , dimer , ligand (biochemistry) , biochemistry , enzyme , organic chemistry , receptor
Citrate synthase (CS) plays a central metabolic role in aerobes and many other organisms. The CS reaction comprises two half‐reactions: a Claisen aldol condensation of acetyl‐CoA (AcCoA) and oxaloacetate (OAA) that forms citryl‐CoA (CitCoA), and CitCoA hydrolysis. Protein conformational changes that `close' the active site play an important role in the assembly of a catalytically competent condensation active site. CS from the thermoacidophile Thermoplasma acidophilum (TpCS) possesses an endogenous Trp fluorophore that can be used to monitor the condensation reaction. The 2.2 Å resolution crystal structure of TpCS fused to a C‐terminal hexahistidine tag (TpCSH6) reported here is an `open' structure that, when compared with several liganded TpCS structures, helps to define a complete path for active‐site closure. One active site in each dimer binds a neighboring His tag, the first nonsubstrate ligand known to occupy both the AcCoA and OAA binding sites. Solution data collectively suggest that this fortuitous interaction is stabilized by the crystalline lattice. As a polar but almost neutral ligand, the active site–tail interaction provides a new starting point for the design of bisubstrate‐analog inhibitors of CS.