title: Structural and functional analysis of PucM, a hydrolase in the ureide pathway and a member of the transthyretin-related protein family.
abstract: The ureide pathway, which produces ureides from uric acid, is an essential purine catabolic process for storing and transporting the nitrogen fixed in leguminous plants and some bacteria. PucM from Bacillus subtilis was recently characterized and found to catalyze the second reaction of the pathway, hydrolyzing 5-hydroxyisourate (HIU), a product of uricase in the first step. PucM has 121 amino acid residues and shows high sequence similarity to the functionally unrelated protein transthyretin (TTR), a thyroid hormone-binding protein. Therefore, PucM belongs to the TTR-related proteins (TRP) family. The crystal structures of PucM at 2.0 A and its complexes with the substrate analogs 8-azaxanthine and 5,6-diaminouracil reveal that even with their overall structure similarity, homotetrameric PucM and TTR are completely different, both in their electrostatic potential and in the size of the active sites located at the dimeric interface. Nevertheless, the absolutely conserved residues across the TRP family, including His-14, Arg-49, His-105, and the C-terminal Tyr-118-Arg-119-Gly-120-Ser-121, indeed form the active site of PucM. Based on the results of site-directed mutagenesis of these residues, we propose a possible mechanism for HIU hydrolysis. The PucM structure determined for the TRP family leads to the conclusion that diverse members of the TRP family would function similarly to PucM as HIU hydrolase.
authors.affiliation: School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-921, Korea.
authors.forename: Du-Kyo
authors.initials: DK
authors.lastname: Jung
authors.forename: Youra
authors.initials: Y
authors.lastname: Lee
authors.forename: Sung Goo
authors.initials: SG
authors.lastname: Park
authors.forename: Byoung Chul
authors.initials: BC
authors.lastname: Park
authors.forename: Ghyung-Hwa
authors.initials: GH
authors.lastname: Kim
authors.forename: Sangkee
authors.initials: S
authors.lastname: Rhee
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D014498:Uracil / Q000031:analogs & derivatives / Q000737:chemistry; D014527:Uric Acid / Q000031:analogs & derivatives / Q000378:metabolism; D014970:Xanthines / Q000737:chemistry
chemical_list: C510200:5-hydroxyisourate; D001426:Bacterial Proteins; D014970:Xanthines; C018221:8-azaxanthine; D014527:Uric Acid; C061549:5,6-diamino-2,4-dihydroxypyrimidine; D014498:Uracil; D006867:Hydrolases