title: Improvement of diffraction quality upon rehydration of dehydrated icosahedral Enterococcus faecalis pyruvate dehydrogenase core crystals.
abstract: Members of the family of 2-oxoacid dehydrogenase multienzyme complexes catalyze the oxidative decarboxylation of alpha-keto acids and are among the most remarkable enzymatic machineries in the living cell. These multienzyme complexes combine a highly symmetric (cubic or icosahedral) core with a dynamic and flexible arrangement of numerous subunits and domains surrounding the core. The center of the complex is formed by either 24 or 60 copies of dihydrolipoamide acetyltransferase (E2)-a multidomain enzyme. The hollow icosahedral cores are composed of 60 identical subunits of the catalytic domain of E2 with a molecular weight of about 1.8 million Da. Bipyramidal crystals suitable for X-ray diffraction of the icosahedral core of the pyruvate dehydrogenase multienzyme complex from Enterococcus faecalis were grown up to 0.7 mm in each dimension. The crystals belong to space group R32 with a = b = 244.3 A (hexagonal setting), and have a solvent content of 73%. The asymmetric unit contains one-third of the molecule, i.e., 20 of the 60 subunits. Initial X-ray crystallographic data to 7 A resolution were collected at cryotemperatures at synchrotron facilities. Interestingly, the diffraction was improved significantly upon rehydrating dehydrated crystals and extended to 4.2 A.
authors.affiliation: Howard Hughes Medical Institute, University of Washington, Seattle 98195-7742, USA.
authors.forename: T
authors.initials: T
authors.lastname: Izard
authors.forename: S
authors.initials: S
authors.lastname: Sarfaty
authors.forename: A
authors.initials: A
authors.lastname: Westphal
authors.forename: A
authors.initials: A
authors.lastname: de Kok
authors.forename: W G
authors.initials: WG
authors.lastname: Hol
mesh_terms: D018360:Crystallography, X-Ray; D013293:Enterococcus faecalis / Q000201:enzymology*; D011768:Pyruvate Dehydrogenase Complex / Q000737:chemistry*; D014867:Water / Q000737:chemistry
chemical_list: D011768:Pyruvate Dehydrogenase Complex; D014867:Water