3M0H
Crystal structure of Pseudomonas stutzeri L-rhamnose isomerase mutant S329F in complex with L-rhamnose
Summary for 3M0H
| Entry DOI | 10.2210/pdb3m0h/pdb |
| Related | 2HCV 2I56 2I57 3M0L 3M0M 3M0V 3M0X 3M0Y |
| Descriptor | L-rhamnose isomerase, MANGANESE (II) ION, L-RHAMNOSE, ... (4 entities in total) |
| Functional Keywords | beta/alpha barrel, homo-tetramer, metal-binding protein, tim barrel, isomerase |
| Biological source | Pseudomonas stutzeri (Pseudomonas perfectomarina) |
| Total number of polymer chains | 4 |
| Total formula weight | 193354.96 |
| Authors | Yoshida, H.,Takeda, K.,Izumori, K.,Kamitori, S. (deposition date: 2010-03-03, release date: 2010-11-10, Last modification date: 2023-11-01) |
| Primary citation | Yoshida, H.,Takeda, K.,Izumori, K.,Kamitori, S. Elucidation of the role of Ser329 and the C-terminal region in the catalytic activity of Pseudomonas stutzeri L-rhamnose isomerase Protein Eng.Des.Sel., 23:919-927, 2010 Cited by PubMed Abstract: Pseudomonas stutzeri l-rhamnose isomerase (l-RhI) is capable of catalyzing the isomerization between various aldoses and ketoses, showing high catalytic activity with broad substrate-specificity compared with Escherichia coli l-RhI. In a previous study, the crystal structure of P. stutzeri l-RhI revealed an active site comparable with that of E. coli l-RhI and d-xylose isomerases (d-XIs) with structurally conserved amino acids, but also with a different residue seemingly responsible for the specificity of P. stutzeri l-RhI, though the residue itself does not interact with the bound substrate. This residue, Ser329, corresponds to Phe336 in E. coli l-RhI and Lys294 in Actinoplanes missouriensis d-XI. To elucidate the role of Ser329 in P. stutzeri l-RhI, we constructed mutants, S329F (E. coli l-RhI type), S329K (A. missouriensis d-XI type), S329L and S329A. Analyses of the catalytic activity and crystal structure of the mutants revealed a hydroxyl group of Ser329 to be crucial for catalytic activity via interaction with a water molecule. In addition, in complexes with substrate, the mutants S329F and S329L exhibited significant electron density in the C-terminal region not observed in the wild-type P. stutzeri l-RhI. The C-terminal region of P. stutzeri l-RhI has flexibility and shows a flip-flop movement at the inter-molecular surface of the dimeric form. PubMed: 20977999DOI: 10.1093/protein/gzq077 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.58 Å) |
Structure validation
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