title: Probing conformational states of glutaryl-CoA dehydrogenase by fragment screening.
abstract: Glutaric acidemia type 1 is an inherited metabolic disorder which can cause macrocephaly, muscular rigidity, spastic paralysis and other progressive movement disorders in humans. The defects in glutaryl-CoA dehydrogenase (GCDH) associated with this disease are thought to increase holoenzyme instability and reduce cofactor binding. Here, the first structural analysis of a GCDH enzyme in the absence of the cofactor flavin adenine dinucleotide (FAD) is reported. The apo structure of GCDH from Burkholderia pseudomallei reveals a loss of secondary structure and increased disorder in the FAD-binding pocket relative to the ternary complex of the highly homologous human GCDH. After conducting a fragment-based screen, four small molecules were identified which bind to GCDH from B. pseudomallei. Complex structures were determined for these fragments, which cause backbone and side-chain perturbations to key active-site residues. Structural insights from this investigation highlight differences from apo GCDH and the utility of small-molecular fragments as chemical probes for capturing alternative conformational states of preformed protein crystals.
authors: Seattle Structural Genomics Center for Infectious Disease (http://www.ssgcid.org), USA. dbegley@embios.com
authors: Darren W
authors: DW
authors: Begley
authors: Douglas R
authors: DR
authors: Davies
authors: Robert C
authors: RC
authors: Hartley
authors: Stephen N
authors: SN
authors: Hewitt
authors: Amanda L
authors: AL
authors: Rychel
authors: Peter J
authors: PJ
authors: Myler
authors: Wesley C
authors: WC
authors: Van Voorhis
authors: Bart L
authors: BL
authors: Staker
authors: Lance J
authors: LJ
authors: Stewart
mesh_terms: D001051:Apoenzymes / Q000737:chemistry; D016957:Burkholderia pseudomallei / Q000201:enzymology*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D050770:Glutaryl-CoA Dehydrogenase / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D010802:Phylogeny; D020836:Protein Structure, Quaternary; D040681:Structural Homology, Protein
chemical_list: D001051:Apoenzymes; D050770:Glutaryl-CoA Dehydrogenase