7JNO
X-ray crystallographic structure of the NS3 helicase domain from Tick-borne encephalitis virus
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | NSLS-II BEAMLINE 17-ID-1 |
| Synchrotron site | NSLS-II |
| Beamline | 17-ID-1 |
| Temperature [K] | 100 |
| Detector technology | PIXEL |
| Collection date | 2019-09-06 |
| Detector | DECTRIS EIGER X 9M |
| Wavelength(s) | 0.920089 |
| Spacegroup name | P 1 21 1 |
| Unit cell lengths | 42.787, 55.442, 91.267 |
| Unit cell angles | 90.00, 93.75, 90.00 |
Refinement procedure
| Resolution | 45.550 - 1.950 |
| R-factor | 0.1841 |
| Rwork | 0.182 |
| R-free | 0.22370 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 5gjc |
| RMSD bond length | 0.010 |
| RMSD bond angle | 1.632 |
| Data reduction software | xia2 (0.6.467-geb301d90-dials-2.2) |
| Data scaling software | Aimless (0.7.4) |
| Phasing software | PHASER |
| Refinement software | REFMAC (5.8.0258) |
Data quality characteristics
| Overall | Inner shell | Outer shell | |
| Low resolution limit [Å] | 45.550 | 45.550 | 2.000 |
| High resolution limit [Å] | 1.950 | 8.720 | 1.950 |
| Rmerge | 0.123 | 0.064 | 0.280 |
| Rmeas | 0.135 | 0.069 | 0.327 |
| Rpim | 0.053 | 0.026 | 0.162 |
| Total number of observations | 190446 | 2540 | 8922 |
| Number of reflections | 31169 | 378 | 2117 |
| <I/σ(I)> | 7.7 | 13.6 | 2.7 |
| Completeness [%] | 99.5 | 99.3 | 93.7 |
| Redundancy | 6.1 | 6.7 | 4.2 |
| CC(1/2) | 0.986 | 0.996 | 0.453 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 293 | Crystals were initially identified in the Morpheus II screen condition G5 (15% PEG 3K, 20% 1,2,4 butanetriol, 1% NDSB-256, 20 mM arginine, 20 mM threonine, 20 mM histidine, 20 mM hydroxylysine, 20 mM trans-4-hydroxy-L-proline, and buffered by 50 mM BES and 50 mM triethanolamine at pH 7.5). Seeds were generated from this condition and used for random microseed matrix screening (rMMS) using a protein concentration of 10-12 mg/ml. Final crystals were identified in the Index HT screen condition H2 (0.2 M Potassium sodium tartrate tetrahydrate, 20% w/v Polyethylene glycol 3,350). Crystals were cryoprotected by transfer to a drop containing a 4:1 ratio of precipitant ethylene glycol |
