title: Cadmium(II) inhibition of human uracil-DNA glycosylase by catalytic water supplantation.
abstract: Toxic metals are known to inhibit DNA repair but the underlying mechanisms of inhibition are still not fully understood. DNA repair enzymes such as human uracil-DNA glycosylase (hUNG) perform the initial step in the base excision repair (BER) pathway. In this work, we showed that cadmium [Cd(II)], a known human carcinogen, inhibited all activity of hUNG at 100 μM. Computational analyses based on 2 μs equilibrium, 1.6 μs steered molecular dynamics (SMD), and QM/MM MD determined that Cd(II) ions entered the enzyme active site and formed close contacts with both D145 and H148, effectively replacing the catalytic water normally found in this position. Geometry refinement by density functional theory (DFT) calculations showed that Cd(II) formed a tetrahedral structure with D145, P146, H148, and one water molecule. This work for the first time reports Cd(II) inhibition of hUNG which was due to replacement of the catalytic water by binding the active site D145 and H148 residues. Comparison of the proposed metal binding site to existing structural data showed that D145:H148 followed a general metal binding motif favored by Cd(II). The identified motif offered structural insights into metal inhibition of other DNA repair enzymes and glycosylases.
authors.affiliation: Department of Chemistry and Biochemistry, San Francisco State University, 1600 Holloway Ave, San Francisco, CA 94132, USA.
authors.forename: Trevor
authors.initials: T
authors.lastname: Gokey
authors.affiliation: Biological Systems and Engineering, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Hang
authors.affiliation: Department of Chemistry and Biochemistry, San Francisco State University, 1600 Holloway Ave, San Francisco, CA 94132, USA.
authors.forename: Anton B
authors.initials: AB
authors.lastname: Guliaev
mesh_terms: D002104:Cadmium / Q000633:toxicity*; D020134:Catalytic Domain / Q000187:drug effects; D019295:Computational Biology / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation; D051981:Uracil-DNA Glycosidase / Q000037:antagonists & inhibitors*; D014867:Water / Q000737:chemistry*
chemical_list: D002104:Cadmium; D014867:Water; D051981:Uracil-DNA Glycosidase