title: Structure- and function-based characterization of a new phosphoglycolate phosphatase from Thermoplasma acidophilum.
abstract: The protein TA0175 has a large number of sequence homologues, most of which are annotated as unknown and a few as belonging to the haloacid dehalogenase superfamily, but has no known biological function. Using a combination of amino acid sequence analysis, three-dimensional crystal structure information, and kinetic analysis, we have characterized TA0175 as phosphoglycolate phosphatase from Thermoplasma acidophilum. The crystal structure of TA0175 revealed two distinct domains, a larger core domain and a smaller cap domain. The large domain is composed of a centrally located five-stranded parallel beta-sheet with strand order S10, S9, S8, S1, S2 and a small beta-hairpin, strands S3 and S4. This central sheet is flanked by a set of three alpha-helices on one side and two helices on the other. The smaller domain is composed of an open faced beta-sandwich represented by three antiparallel beta-strands, S5, S6, and S7, flanked by two oppositely oriented alpha-helices, H3 and H4. The topology of the large domain is conserved; however, structural variation is observed in the smaller domain among the different functional classes of the haloacid dehalogenase superfamily. Enzymatic assays on TA0175 revealed that this enzyme catalyzed the dephosphorylation of phosphoglycolate in vitro with similar kinetic properties seen for eukaryotic phosphoglycolate phosphatase. Activation by divalent cations, especially Mg2+, and competitive inhibition behavior with Cl- ions are similar between TA0175 and phosphoglycolate phosphatase. The experimental evidence presented for TA0175 is indicative of phosphoglycolate phosphatase.
authors.affiliation: Argonne National Laboratory, Structural Biology Center, Argonne, Illinois 60439, USA.
authors.forename: Youngchang
authors.initials: Y
authors.lastname: Kim
authors.forename: Alexander F
authors.initials: AF
authors.lastname: Yakunin
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Kuznetsova
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Xu
authors.forename: Micha
authors.initials: M
authors.lastname: Pennycooke
authors.forename: Jun
authors.initials: J
authors.lastname: Gu
authors.forename: Fred
authors.initials: F
authors.lastname: Cheung
authors.forename: Michael
authors.initials: M
authors.lastname: Proudfoot
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Arrowsmith
authors.forename: Andrzej
authors.initials: A
authors.lastname: Joachimiak
authors.forename: Aled M
authors.initials: AM
authors.lastname: Edwards
authors.forename: Dinesh
authors.initials: D
authors.lastname: Christendat
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D018724:Spinacia oleracea / Q000201:enzymology; D013822:Thermoplasma / Q000201:enzymology*
chemical_list: D019843:Archaeal Proteins; C022408:phosphoglycolate phosphatase; D010744:Phosphoric Monoester Hydrolases