1tq0
|
Crystal structure of the potent anticoagulant thrombin mutant W215A/E217A in free form
OverviewOverview
The thrombin mutant W215A/E217A features a drastically impaired catalytic, activity toward chromogenic and natural substrates but efficiently, activates the anticoagulant protein C in the presence of thrombomodulin., As the remarkable anticoagulant properties of this mutant continue to be, unraveled in preclinical studies, we solved the x-ray crystal structures, of its free form and its complex with the active site inhibitor, H-d-Phe-Pro-Arg-CH(2)Cl (PPACK). The PPACK-bound structure of W215A/E217A, is identical to the structure of the PPACK-bound slow form of thrombin. On, the other hand, the structure of the free form reveals a collapse of the, 215-217 strand that crushes the primary specificity pocket. The collapse, results from abrogation of the stacking interaction between Phe-227 and, Trp-215 and the polar interactions of Glu-217 with Thr-172 and Lys-224., Other notable changes are a rotation of the carboxylate group of Asp-189, breakage of the H-bond between the catalytic residues Ser-195 and His-57, breakage of the ion pair between Asp-222 and Arg-187, and significant, disorder in the 186- and 220-loops that define the Na(+) site. These, findings explain the impaired catalytic activity of W215A/E217A and, demonstrate that the analysis of the molecular basis of substrate, recognition by thrombin and other proteases requires crystallization of, both the free and bound forms of the enzyme.
DiseaseDisease
Known diseases associated with this structure: Dysprothrombinemia OMIM:[176930], Hyperprothrombinemia OMIM:[176930], Hypoprothrombinemia OMIM:[176930]
About this StructureAbout this Structure
1TQ0 is a Protein complex structure of sequences from Homo sapiens. Active as Thrombin, with EC number 3.4.21.5 Full crystallographic information is available from OCA.
ReferenceReference
The anticoagulant thrombin mutant W215A/E217A has a collapsed primary specificity pocket., Pineda AO, Chen ZW, Caccia S, Cantwell AM, Savvides SN, Waksman G, Mathews FS, Di Cera E, J Biol Chem. 2004 Sep 17;279(38):39824-8. Epub 2004 Jul 13. PMID:15252033
Page seeded by OCA on Mon Nov 12 19:26:52 2007