Inhibition of Trypsin and Thrombin by Amino(4-amidinophenyl)methanephosphonate Diphenyl Ester Derivatives: X-ray Structures and Molecular Models
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- 作者：Jay A. Bertrand ; Josef Oleksyszyn ; Chih-Min Kam ; Bogdan Boduszek ; Steven Presnell ; R. Richard Plaskon ; F. L. Suddath ; James C. Powers ; and Loren Dean Williams
- 刊名：Biochemistry
- 出版年：1996
- 出版时间：March 12, 1996
- 年：1996
- 卷：35
- 期：10
- 页码：3147 - 3155
- 全文大小：451K
- 年卷期：v.35,no.10(March 12, 1996)
- ISSN：1520-4995
文摘
X-ray structures of trypsin from bovine pancreas inactivated bydiphenyl [N-(benzyloxycarbonyl)amino](4-amidinophenyl)methanephosphonate[Z-(4-AmPhGly)P(OPh)2] were determinedat 113and 293 K to 1.8 Å resolution and refined to R factors of0.211 (113 K) and 0.178 (293 K). The structuresreveal a tetrahedral phosphorus covalently bonded to the O
of theactive site serine. Covalent bondformation is accompanied by the loss of both phenoxy groups. TheD-stereoisomer ofZ-(4-AmPhGly)P(OPh)2 is not observed in the complex. TheL-stereoisomer of the inhibitor forms contacts withseveralresidues in the trypsin active site. One of the phosphonateoxygens is inserted into the oxyanion hole andforms hydrogen bonds to the amides of Gly193, Asp194, and Ser195.The second phosphonate oxygenforms hydrogen bonds to N
2 of His 57. Thep-amidinophenylglycine moiety binds into thetrypsinprimary specificity pocket, interacting with Asp189. The amideforms a hydrogen bond to the carbonyloxygen atom of Ser214. The inhibitor moiety, from the 113 Kstructure of trypsin inactivated by thereaction product of Z-(4-AmPhGly)P(OPh)2,was docked into human thrombin [Bode, W., Mayr, I.,Baumann, U., Huber, R., Stone, S. R., & Hofsteenge, J. (1989)EMBO J. 8, 3467-3475] and energyminimized. The inhibitor fits well into the thrombin active site,forming favorable contacts similar tothose in the trypsin complex with no bad contacts.
of theactive site serine. Covalent bondformation is accompanied by the loss of both phenoxy groups. TheD-stereoisomer ofZ-(4-AmPhGly)P(OPh)2 is not observed in the complex. TheL-stereoisomer of the inhibitor forms contacts withseveralresidues in the trypsin active site. One of the phosphonateoxygens is inserted into the oxyanion hole andforms hydrogen bonds to the amides of Gly193, Asp194, and Ser195.The second phosphonate oxygenforms hydrogen bonds to N2 of His 57. Thep-amidinophenylglycine moiety binds into thetrypsinprimary specificity pocket, interacting with Asp189. The amideforms a hydrogen bond to the carbonyloxygen atom of Ser214. The inhibitor moiety, from the 113 Kstructure of trypsin inactivated by thereaction product of Z-(4-AmPhGly)P(OPh)2,was docked into human thrombin [Bode, W., Mayr, I.,Baumann, U., Huber, R., Stone, S. R., & Hofsteenge, J. (1989)EMBO J. 8, 3467-3475] and energyminimized. The inhibitor fits well into the thrombin active site,forming favorable contacts similar tothose in the trypsin complex with no bad contacts.