Crystal structure of chloromuconate cycloisomerase from Alcaligenes eutrophus JMP134 (pJP4) at 3 Å resolution

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 50; no. 1; pp. 75 - 84
• Main Authors: Hoier, H., Schlömann, M., Hammer, A., Glusker, J. P., Carrell, H. L., Goldman, A., Stezowski, J. J., Heinemann, U.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.01.1994
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S090744499300900X

Chloromuconate cycloisomerase (E.C. 5.5.1.7) is an enzyme involved in the 2,4‐dichlorophenoxyacetate degradation pathway of Alcaligenes eutrophus JMP134 (pJP4). The crystal structure of this protein was determined at 3 Å resolution by molecular‐replacement techniques using atomic coordinates from the reported crystal structure of the homologous muconate cycloisomerase (E.C. 5.5.1.1) from Pseudomonas putida as the search model (42% identical positions in the sequences). Structure refinement by simulated‐annealing and restrained least‐squares techniques converged at R = 0.195. In the crystals studied, space group I4, the protein is present as two octamers per unit cell with two subunits per asymmetric unit. Each subunit consists of two globular domains, one of which forms an α/β‐barrel. Comparison of this structure with that of muconate cycloisomerase reveals the reasons for the altered substrate specificity of chloromuconate cycloisomerase. Marked differences are observed in polarity, accessibility and hydrogen‐bonding potential in the channel leading into the active site as well as in the active center itself.