TY - JOUR
T1 - Crystal structure of constitutive endothelial nitric oxide synthase
T2 - A paradigm for pterin function involving a novel metal center
AU - Raman, C. S.
AU - Li, Huiying
AU - Martásek, Pavel
AU - Král, Vladimir
AU - Masters, Bettie Sue S.
AU - Poulos, Thomas L.
N1 - Funding Information:
C. S. R. is delighted to acknowledge W. Lipscomb and B. Vallee for stimulating dialogs on zinc; E. de La Fortelle for constructive suggestions regarding SHARP phasing; C. Nielsen and X-P. Dai for help with CCD data processing; L. Sanchez for ENDHKL; T. Terwilliger for SOLVE; S. Islam for PREPI; A. Greenwood for help with graphics; M. Connolly for MSP; S. Jain, T. Rydel, D. Schuller, W. Sessa, I. Sevrioukova, and S. Veeraraghavan for helpful discussions; M. Sundaramoorthy for stimulating discussions and help with SHELXL refinement; G. Petsko for EMP; and R. Stevens for generously providing coordinates of pterin-bound TyrOH before publication. We are grateful to A. McPherson for the use of his laboratory facilities and advice. We would like to thank the following synchrotron sources for timely access to their beamlines and their staffs for help with data collection: ALS (T. Earnest and L-W. Hung, 5.0.2); CHESS (A. Deacon, D. Thiel and staffs of F-1 and F-2); NSLS (M. Capel, X12B); and SSRL (H. Bellamy, 1-5; A. Cohen, P. Ellis, P. Kuhn, H-D. Nuhn, and M. Soltis, 7-1 and 9-1). This work was supported by the National Institutes of Health (grants GM32688 and GM57353 to T. L. P.; HL30050 and GM52419 to B. S. S. M.), the Robert A. Welch Foundation (grant AQ-1192 to B. S. S. M.), the Ministry of Education of Czech Republic (V. K.), the Howard Hughes Medical Institute (V. K.), and Berlex Corp. (T. L. P.). C. S. R. is a Fellow of the American Heart Association (Western States Affiliate) and is grateful for their support of this work.
PY - 1998/12/23
Y1 - 1998/12/23
N2 - Nitric oxide, a key signaling molecule, is produced by a family of enzymes collectively called nitric oxide synthases (NOS). Here, we report the crystal structure of the heme domain of endothelial NOS in tetrahydrobiopterin (H4B)-free and -bound forms at 1.95 Å and 1.9 Å resolution, respectively. In both structures a zinc ion is tetrahedrally coordinated to pairs of symmetry-related cysteine residues at the dimer interface. The phylogenetically conserved Cys-(X)4-Cys motif and its strategic location establish a structural role for the metal center in maintaining the integrity of the H4B-binding site. The unexpected recognition of the substrate, L-arginine, at the H4B site indicates that this site is poised to stabilize a positively charged pterin ring and suggests a model involving a cationic pterin radical in the catalytic cycle.
AB - Nitric oxide, a key signaling molecule, is produced by a family of enzymes collectively called nitric oxide synthases (NOS). Here, we report the crystal structure of the heme domain of endothelial NOS in tetrahydrobiopterin (H4B)-free and -bound forms at 1.95 Å and 1.9 Å resolution, respectively. In both structures a zinc ion is tetrahedrally coordinated to pairs of symmetry-related cysteine residues at the dimer interface. The phylogenetically conserved Cys-(X)4-Cys motif and its strategic location establish a structural role for the metal center in maintaining the integrity of the H4B-binding site. The unexpected recognition of the substrate, L-arginine, at the H4B site indicates that this site is poised to stabilize a positively charged pterin ring and suggests a model involving a cationic pterin radical in the catalytic cycle.
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U2 - 10.1016/S0092-8674(00)81718-3
DO - 10.1016/S0092-8674(00)81718-3
M3 - Article
C2 - 9875848
AN - SCOPUS:0032416666
SN - 0092-8674
VL - 95
SP - 939
EP - 950
JO - Cell
JF - Cell
IS - 7
ER -