Structure of N-formimino-L-glutamate iminohydrolase from Pseudomonas aeruginosa. Academic Article uri icon


  • N-Formimino-l-glutamate iminohydrolase (HutF), from Pseudomonas aeruginosa with a locus tag of Pa5106 ( gi|15600299 ), is a member of the amidohydrolase superfamily. This enzyme catalyzes the deamination of N-formimino-l-glutamate to N-formyl-l-glutamate and ammonia in the histidine degradation pathway. The crystal structure of Pa5106 was determined in the presence of the inhibitors N-formimino-l-aspartate and N-guanidino-l-glutaric acid at resolutions of 1.9 and 1.4 , respectively. The structure of an individual subunit is composed of two domains with the larger domain folding as a distorted (/)8-barrel. The (/)8-barrel domain is composed of eight -strands flanked by 11 -helices, whereas the smaller domain is made up of eight -strands. The active site of Pa5106 contains a single zinc atom that is coordinated by His-56, His-58, His-232, and Asp-320. The nucleophilic solvent water molecule coordinates with the zinc atom at a distance of 2.0 and is hydrogen bonded to Asp-320 and His-269. The -carboxylate groups of both inhibitors are hydrogen bonded to the imidazole moiety of His-206, the hydroxyl group of Tyr-121, and the side chain amide group of Gln-61. The side chain carboxylate groups of the two inhibitors are ion-paired with the guanidino groups of Arg-209 and Arg-82. Computational docking of high-energy tetrahedral intermediate forms of the substrate, N-formimino-l-glutamate, to the three-dimensional structure of Pa5106 suggests that this compound likely undergoes a re-faced nucleophilic attack at the formimino group by the metal-bound hydroxide. A catalytic mechanism of the reaction catalyzed by Pa5106 is proposed.

published proceedings

  • Biochemistry

author list (cited authors)

  • Fedorov, A. A., Mart-Arbona, R., Nemmara, V. V., Hitchcock, D., Fedorov, E. V., Almo, S. C., & Raushel, F. M.

citation count

  • 2

complete list of authors

  • Fedorov, Alexander A||Martí-Arbona, Ricardo||Nemmara, Venkatesh V||Hitchcock, Daniel||Fedorov, Elena V||Almo, Steven C||Raushel, Frank M

publication date

  • January 2015