7ASZ
L-2-haloacid dehalogenase H190A mutant from Zobellia galactanivorans
Summary for 7ASZ
| Entry DOI | 10.2210/pdb7asz/pdb |
| Descriptor | (S)-2-haloacid dehalogenase, PHOSPHATE ION, THIOCYANATE ION, ... (4 entities in total) |
| Functional Keywords | haloacid dehalogenase, rossmann fold, cytoplasmic, hydrolase |
| Biological source | Zobellia galactanivorans (strain DSM 12802 / CCUG 47099 / CIP 106680 / NCIMB 13871 / Dsij) |
| Total number of polymer chains | 2 |
| Total formula weight | 52924.75 |
| Authors | Grigorian, E.,Roret, T.,Czjzek, M.,Leblanc, C.,Delage, L. (deposition date: 2020-10-28, release date: 2021-09-08, Last modification date: 2024-01-31) |
| Primary citation | Grigorian, E.,Roret, T.,Czjzek, M.,Leblanc, C.,Delage, L. X-ray structure and mechanism of ZgHAD, a L-2-haloacid dehalogenase from the marine Flavobacterium Zobellia galactanivorans. Protein Sci., :e4540-e4540, 2022 Cited by PubMed Abstract: Haloacid dehalogenases are potentially involved in bioremediation of contaminated environments and few have been biochemically characterized from marine organisms. The l-2-haloacid dehalogenase (l-2-HAD) from the marine Bacteroidetes Zobellia galactanivorans Dsij (ZgHAD) has been shown to catalyze the dehalogenation of C2 and C3 short-chain l-2-haloalkanoic acids. To better understand its catalytic properties, its enzymatic stability, active site, and 3D structure were analyzed. ZgHAD demonstrates high stability to solvents and a conserved catalytic activity when heated up to 60°C, its melting temperature being at 65°C. The X-ray structure of the recombinant enzyme was solved by molecular replacement. The enzyme folds as a homodimer and its active site is very similar to DehRhb, the other known l-2-HAD from a marine Rhodobacteraceae. Marked differences are present in the putative substrate entrance sites of the two enzymes. The H179 amino acid potentially involved in the activation of a catalytic water molecule was confirmed as catalytic amino acid through the production of two inactive site-directed mutants. The crystal packing of 13 dimers in the asymmetric unit of an active-site mutant, ZgHAD-H179N, reveals domain movements of the monomeric subunits relative to each other. The involvement of a catalytic His/Glu dyad and substrate binding amino acids was further confirmed by computational docking. All together our results give new insights into the catalytic mechanism of the group of marine l-2-HAD. PubMed: 36502283DOI: 10.1002/pro.4540 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.88 Å) |
Structure validation
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