Download MendeleyCommunication between the Zinc and Tetrahydrobiopterin Binding Sites in Nitric Oxide Synthase.
Chreifi, G., Li, H., Mcinnes, C.R., Gibson, C.L., Suckling, C.J., Poulos, T.L.(2014) Biochemistry 53: 4216
- PubMed: 24819538
- DOI: https://doi.org/10.1021/bi5003986
- Primary Citation of Related Structures:
4CUL, 4CUM, 4CUN, 4CVG - PubMed Abstract:
The nitric oxide synthase (NOS) dimer is stabilized by a Zn(2+) ion coordinated to four symmetry-related Cys residues exactly along the dimer 2-fold axis. Each of the two essential tetrahydrobiopterin (H4B) molecules in the dimer interacts directly with the heme, and each H4B molecule is ~15 ? from the Zn(2+). We have determined the crystal structures of the bovine endothelial NOS dimer oxygenase domain bound to three different pterin analogues, which reveal an intimate structural communication between the H4B and Zn(2+) sites. The binding of one of these compounds, 6-acetyl-2-amino-7,7-dimethyl-7,8-dihydro-4(3H)-pteridinone (1), to the pterin site and Zn(2+) binding are mutually exclusive. Compound 1 both directly and indirectly disrupts hydrogen bonding between key residues in the Zn(2+) binding motif, resulting in destabilization of the dimer and a complete disruption of the Zn(2+) site. Addition of excess Zn(2+) stabilizes the Zn(2+) site at the expense of weakened binding of 1. The unique structural features of 1 that disrupt the dimer interface are extra methyl groups that extend into the dimer interface and force a slight opening of the dimer, thus resulting in disruption of the Zn(2+) site. These results illustrate a very delicate balance of forces and structure at the dimer interface that must be maintained to properly form the Zn(2+), pterin, and substrate binding sites.
Organizational Affiliation:
Departments of ?Molecular Biology and Biochemistry, ?Chemistry, and ¡ìPharmaceutical Sciences, University of California , Irvine, California 92697-3900, United States.
