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