| US 7,590,494 B1 | ||
| Drug design based on the structure of LTA4 hydrolase | ||
| Jesper Z. Haeggström, Valhallavägen 145, SE-115 31 Stockholm (Sweden); Pär Nordlund, Gruvbacken 2, SE-116 34 Stockholm (Sweden); and Marjolein Thunissen, Svinningevägen 26, SE-184 92 Åkersberga (Sweden) | ||
| Appl. No. 9/914,451 PCT Filed Feb. 28, 2000, PCT No. PCT/SE00/00384 § 371(c)(1), (2), (4) Date Dec. 20, 2001, PCT Pub. No. WO00/50577, PCT Pub. Date Aug. 31, 2000. |
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| Claims priority of provisional application 60/122110, filed on Feb. 26, 1999. | ||
| Claims priority of application No. 9900722 (SE), filed on Feb. 26, 1999. | ||
| Int. Cl. G01N 31/00 (2006.01); C12N 9/14 (2006.01); G06G 7/58 (2006.01) | ||
| U.S. Cl. 702—27 [435/195; 703/11] | 7 Claims |
| 1. A method of identifying compounds that bind to a leukotriene A4 (LTA4) hydrolase comprising the amino acid sequence of SEQ ID NO: 1, the method comprising the steps of:
(a) crystallizing a purified LTA4 hydrolase in the presence of bestatin to form a co-crystal of LTA4 hydrolase and bestatin, wherein crystallization is performed by liquid-liquid diffusion in a capillary using equal volumes
of a buffer:
enzyme solution consisting of:i) a buffer solution consisting of 28% PEG8000, 0.1 M Na-acetate, 0.1 M imidazole at a pH of 6.8 and with 5 mM YbCl3 as an additive; andii) an enzyme solution consisting of 5 mg/ml LTA4 hydrolase comprising the amino acid sequence of SEQ ID NO:1 in 10 mM Tris-HCl at a pH of 8, supplemented with 1 mM bestatin;wherein the crystallization results in a LTA4 hydrolase crystal having the space group P21212 and the unit cell dimensions a=67.59 Å, b=133.51 Å, and c=83.40 Å and wherein α=β=γ=90°;
(b) determining the atomic coordinates of LTA4 hydrolase from the co-crystal obtained in step (a); and
(c) screening the atomic coordinates of a set of candidate compounds against the atomic coordinates obtained in step (b) to
identify compounds that bind to the LTA4 hydrolase.
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