title: The crystal structure and active site location of isocitrate lyase from the fungus Aspergillus nidulans.
abstract: BACKGROUND
  Isocitrate lyase catalyses the first committed step of the carbon-conserving glyoxylate bypass, the Mg(2+)-dependent reversible cleavage of isocitrate into succinate and glyoxylate. This metabolic pathway is an inviting target for the control of a number of diseases, because the enzymes involved in this cycle have been identified in many pathogens including Mycobacterium leprae and Leishmania.
  
  
  RESULTS
  As part of a programme of rational drug design the structure of the tetrameric Aspergillus nidulans isocitrate lyase and its complex with glyoxylate and a divalent cation have been solved to 2.8 A resolution using X-ray diffraction. Each subunit comprises two domains, one of which adopts a folding pattern highly reminiscent of the triose phosphate isomerase (TIM) barrel. A 'knot' between subunits observed in the three-dimensional structure, involving residues towards the C terminus, implies that tetramer assembly involves considerable flexibility in this part of the protein.
  
  
  CONCLUSIONS
  Difference Fourier analysis together with the pattern of sequence conservation has led to the identification of both the glyoxylate and metal binding sites and implicates the C-terminal end of the TIM barrel as the active site, which is consistent with studies of other enzymes with this fold. Two disordered regions of the polypeptide chain lie close to the active site, one of which includes a critical cysteine residue suggesting that conformational rearrangements are essential for catalysis. Structural similarities between isocitrate lyase and both PEP mutase and enzymes belonging to the enolase superfamily suggest possible relationships in aspects of the mechanism.
authors.affiliation: Department of Molecular Biology and Biotechnology, Krebs Institute for Biomolecular Research, The University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: K
authors.initials: K
authors.lastname: Britton
authors.forename: S
authors.initials: S
authors.lastname: Langridge
authors.forename: P J
authors.initials: PJ
authors.lastname: Baker
authors.forename: K
authors.initials: K
authors.lastname: Weeradechapon
authors.forename: S E
authors.initials: SE
authors.lastname: Sedelnikova
authors.forename: J R
authors.initials: JR
authors.lastname: De Lucas
authors.forename: D W
authors.initials: DW
authors.lastname: Rice
authors.forename: G
authors.initials: G
authors.lastname: Turner
mesh_terms: D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000201:enzymology*; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D007522:Isocitrate Lyase / Q000737:chemistry* / Q000378:metabolism*; D008670:Metals / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017875:Phosphotransferases (Phosphomutases) / Q000737:chemistry; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
chemical_list: D008670:Metals; D007522:Isocitrate Lyase; D017875:Phosphotransferases (Phosphomutases); C057167:phosphoenolpyruvate mutase