title: Identification of the structural similarity in the functionally related amidohydrolases acting on the cyclic amide ring.
abstract: The functionally related amidohydrolases, including D-hydantoinases, dihydropyrimidinases, allantoinases and dihydro-orotases, share a similar catalytic function of acting on the cyclic amide ring. We aligned 16 amidohydrolases by taking account of the conservative substitution and found a number of highly conserved regions and invariant amino acid residues. Analyses of the secondary structure and hydropathy profile of the enzymes revealed a significant degree of similarity in the conserved regions. Among the regions, the long stretched region I is of particular interest, because it is mainly composed of invariant amino acid residues, showing a similarity of 69% for the enzymes. A search of the protein data bank using the sequence of the conserved region I identified a number of proteins possessing a similar catalytic property, providing a clue that this region might be linked with the catalytic function. As a particular sequence, one aspartic acid and four histidine residues are found to be rigidly conserved in the functionally related amidohydrolases. In order to investigate the significance of the conserved residues, site-directed mutagenesis was carried out typically for the D-hydantoinase gene cloned from Bacillus stearothermophilus SD1. These residues were found to be essential for metal binding as well as catalysis, strongly implying that these invariant residues play a critical role in other enzymes as well as in D-hydantoinase. On the basis of the similar catalytic function and existence of the rigidly conserved sequence, we propose a close evolutionary relationship among the functionally related amido hydrolases, including D-hydantoinase, dihydropyrimidinase, allantoinase and dihydroorotase.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon, South Korea.
authors.forename: G J
authors.initials: GJ
authors.lastname: Kim
authors.forename: H S
authors.initials: HS
authors.lastname: Kim
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D004080:Dihydroorotase / Q000737:chemistry; D001411:Geobacillus stearothermophilus; D006639:Histidine / Q000737:chemistry; D008667:Metalloproteins / Q000737:chemistry; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
chemical_list: D001426:Bacterial Proteins; D008667:Metalloproteins; D011994:Recombinant Proteins; D006639:Histidine; D000581:Amidohydrolases; C020047:dihydropyrimidinase; D004080:Dihydroorotase; C019659:allantoinase