1TUW
Structural and Functional Analysis of Tetracenomycin F2 Cyclase from Streptomyces glaucescens: A Type-II Polyketide Cyclase
Experimental procedure
| Experimental method | MAD |
| Source type | ROTATING ANODE |
| Source details | RIGAKU RU200 |
| Temperature [K] | 273 |
| Detector technology | AREA DETECTOR |
| Collection date | 2000-03-01 |
| Detector | SIEMENS HI-STAR |
| Spacegroup name | P 65 2 2 |
| Unit cell lengths | 46.700, 46.700, 188.300 |
| Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
| Resolution | 30.000 - 1.900 |
| R-factor | 0.199 |
| Rwork | 0.199 |
| R-free | 0.24500 |
| Structure solution method | MAD |
| RMSD bond length | 0.017 |
| RMSD bond angle | 2.610 |
| Data reduction software | XDS |
| Data scaling software | XDS |
| Phasing software | SOLVE |
| Refinement software | TNT |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 30.000 | 2.000 |
| High resolution limit [Å] | 1.900 | 1.900 |
| Rmerge | 0.027 | 0.088 |
| Number of reflections | 11355 | |
| <I/σ(I)> | 23.1 | 5.4 |
| Completeness [%] | 92.0 | 84 |
| Redundancy | 8.8 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | MICROBATCH | 7.5 | 278 | Crystals of tetracenomycin F2 cyclase were grown by microbatch from a solution composed of 2.2 M (NH4)2SO4, 3 mg/ml protein, 50 mM HEPES pH 7.5 at 4 C (11). Large crystals were obtained by macroseeding with small crystals (0.06 mm x 0.08 mm) washed in macroseeding 1.4 M (NH4)2SO4, 50 mM HEPES pH 7.5 to slightly dissolve the crystal surfaces. After 3 to 4 weeks the crystals attained a size of 0.3 x 0.3 x 0.6 mm. Crystals of the selenomethionine labeled protein were obtained in a similar manner., Micro batch, temperature 278K |
