2B9C
Structure of tropomyosin's mid-region: bending and binding sites for actin
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | CHESS BEAMLINE F1 |
Synchrotron site | CHESS |
Beamline | F1 |
Temperature [K] | 100 |
Detector technology | CCD |
Detector | ADSC QUANTUM 4 |
Wavelength(s) | 0.9474 |
Spacegroup name | P 65 |
Unit cell lengths | 80.515, 80.515, 112.453 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 20.000 - 2.300 |
Rwork | 0.256 |
R-free | 0.29600 |
Structure solution method | MAD |
RMSD bond length | 0.051 |
RMSD bond angle | 3.866 |
Data reduction software | DENZO |
Data scaling software | SCALEPACK |
Phasing software | SOLVE (2.03) |
Refinement software | CNS |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 20.000 | 2.380 |
High resolution limit [Å] | 2.300 | 2.300 |
Rmerge | 0.069 | 0.337 |
Number of reflections | 18253 | |
<I/σ(I)> | 15.9 | 4.6 |
Completeness [%] | 99.4 | 100 |
Redundancy | 4.7 | 4.5 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 7 | 289 | Native: Three microliters of protein solution [6.35mg/ml MidTm; 20mM Tris-HCl pH7.0; 100mM NaCl; 2mM 2-mercaptoethanol and 2mM NaN3] were combined with two microliters of 14% PEG2k-MME, and the resultant drop was equilibrated against 1ml reservoir solution [7.5% PEG2k-MME; 120mM NaCl; 24mM Tris-HCl pH7.0] by vapor diffusion at 16 C. Se-Met: Two microliters of protein solution [18mg/ml MidTm; 20mM Tris-HCl pH6.9; 100mM NaCl; 10mM mercaptoethanol and 2mM NaN3] were combined with two microliters of precipitant solution [28-32% PEG2K-MME; 50mM NaCl; 20mM Tris-HCl pH6.9 and 5mM mercaptoethanol]. The resultant mixture was equilibrated against 1ml reservoir solution [24% PEG2K-MME; 50mM NaCl; 20mM Tris-HCL pH6.9 and 5mM mercaptoethanol] by vapor diffusion at 16C. , VAPOR DIFFUSION, HANGING DROP, temperature 289K |
1 | VAPOR DIFFUSION, HANGING DROP | 7 | 289 | Native: Three microliters of protein solution [6.35mg/ml MidTm; 20mM Tris-HCl pH7.0; 100mM NaCl; 2mM 2-mercaptoethanol and 2mM NaN3] were combined with two microliters of 14% PEG2k-MME, and the resultant drop was equilibrated against 1ml reservoir solution [7.5% PEG2k-MME; 120mM NaCl; 24mM Tris-HCl pH7.0] by vapor diffusion at 16 C. Se-Met: Two microliters of protein solution [18mg/ml MidTm; 20mM Tris-HCl pH6.9; 100mM NaCl; 10mM mercaptoethanol and 2mM NaN3] were combined with two microliters of precipitant solution [28-32% PEG2K-MME; 50mM NaCl; 20mM Tris-HCl pH6.9 and 5mM mercaptoethanol]. The resultant mixture was equilibrated against 1ml reservoir solution [24% PEG2K-MME; 50mM NaCl; 20mM Tris-HCL pH6.9 and 5mM mercaptoethanol] by vapor diffusion at 16C. , VAPOR DIFFUSION, HANGING DROP, temperature 289K |
1 | VAPOR DIFFUSION, HANGING DROP | 7 | 289 | Native: Three microliters of protein solution [6.35mg/ml MidTm; 20mM Tris-HCl pH7.0; 100mM NaCl; 2mM 2-mercaptoethanol and 2mM NaN3] were combined with two microliters of 14% PEG2k-MME, and the resultant drop was equilibrated against 1ml reservoir solution [7.5% PEG2k-MME; 120mM NaCl; 24mM Tris-HCl pH7.0] by vapor diffusion at 16 C. Se-Met: Two microliters of protein solution [18mg/ml MidTm; 20mM Tris-HCl pH6.9; 100mM NaCl; 10mM mercaptoethanol and 2mM NaN3] were combined with two microliters of precipitant solution [28-32% PEG2K-MME; 50mM NaCl; 20mM Tris-HCl pH6.9 and 5mM mercaptoethanol]. The resultant mixture was equilibrated against 1ml reservoir solution [24% PEG2K-MME; 50mM NaCl; 20mM Tris-HCL pH6.9 and 5mM mercaptoethanol] by vapor diffusion at 16C. , VAPOR DIFFUSION, HANGING DROP, temperature 289K |