1C1U
RECRUITING ZINC TO MEDIATE POTENT, SPECIFIC INHIBITION OF SERINE PROTEASES
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | ROTATING ANODE |
Temperature [K] | 298 |
Detector technology | IMAGE PLATE |
Collection date | 1998-01-23 |
Detector | RIGAKU RAXIS IV++ |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 71.680, 72.120, 73.110 |
Unit cell angles | 90.00, 101.62, 90.00 |
Refinement procedure
Resolution | 7.500 - 1.750 |
R-factor | 0.206 |
Rwork | 0.206 |
R-free | 0.23500 |
Structure solution method | DIFFERENCE FOURIER PLUS REFINEMENT |
RMSD bond length | 0.018 |
RMSD bond angle | 4.100 |
Data reduction software | bioteX ((MSC)) |
Data scaling software | bioteX |
Phasing software | X-PLOR |
Refinement software | X-PLOR (3.1) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 38.780 | 1.830 |
High resolution limit [Å] | 1.450 | 1.750 |
Rmerge | 0.085 | 0.250 |
Number of reflections | 35066 | |
<I/σ(I)> | 7.8 | 1.8 |
Completeness [%] | 77.0 | 50 |
Redundancy | 1.9 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | unknown * | 8.2 | THROMBIN WAS PURCHASED FROM HAEMATOLOGIC TECHNOLOGIES, INC. AND ACETYL-HIRUDIN FROM BACHEM. THROMBIN WAS PREPARED AS DESCRIBED (SKRZPCZAK-JANKUN ET AL., 1991) .THROMBIN (1.0 MG/ML IN 50 MM HEPES, 50 % GLYCEROL, PH 7.0) WAS INCUBATED WITH 1.0 MM ACETYL-HIRUDIN, 1.0 MM HEMI-BABIM, 1.0 MM ZN+2 FOR 1 HR AT 4 DEG C. GLYCEROL WAS REMOVED AND THE COMPLEX CONCENTRATED WITH A CENTRICON 10 ( AMICON) TO 8.6 MG/ML AS DETERMINED BY THE BIORAD PROTEIN ASSAY KIT USING BOVINE SERUM ALBUMIN. CRYSTALS OF THROMBIN-ACETYL-HIRUDIN-HEMI-BABIM-ZN+2 WERE GROWN IN HANGING DROPS BY VAPOR DIFFUSION AFTER STREAK SEEDING. THE DROPS WERE MADE FROM 5 MICROLITERS OF COMPLEX AND 5 MICROLITERS OF RESERVOIR SOLUTION ( 0.10 M TRIS,0.50 M NACL, 22 % (BY VOLUME) PEG 4K, PH 8.20). |