Catalytic Cycle of the N-Acetylglucosaminidase NagZ from Pseudomonas aeruginosa.

ABSTRACT: The N-acetylglucosaminidase NagZ of Pseudomonas aeruginosa catalyzes the first cytoplasmic step in recycling of muropeptides, cell-wall-derived natural products. This reaction regulates gene expression for the ?-lactam resistance enzyme, ?-lactamase. The enzyme catalyzes hydrolysis of N-acetyl-?-d-glucosamine-(1?4)-1,6-anhydro-N-acetyl-?-d-muramyl-peptide (1) to N-acetyl-?-d-glucosamine (2) and 1,6-anhydro-N-acetyl-?-d-muramyl-peptide (3). The structural and functional aspects of catalysis by NagZ were investigated by a total of seven X-ray structures, three computational models based on the X-ray structures, molecular-dynamics simulations and mutagenesis. The structural insights came from the unbound state and complexes of NagZ with the substrate, products and a mimetic of the transient oxocarbenium species, which were prepared by synthesis. The mechanism involves a histidine as acid/base catalyst, which is unique for glycosidases. The turnover process utilizes covalent modification of D244, requiring two transition-state species and is regulated by coordination with a zinc ion. The analysis provides a seamless continuum for the catalytic cycle, incorporating large motions by four loops that surround the active site.

SUBMITTER: Acebron I 

PROVIDER: S-EPMC6873925 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature