Crystallisation of Wild-Type and Variant Forms of a Recombinant Plant Enzyme β-D-Glucan Glucohydrolase from Barley (Hordeum vulgare L.) and Preliminary X-ray Analysis
Abstract
:1. Introduction
2. Results and Discussion
2.1. Protein Expression and Purification
2.2. Protein Crystallisation
2.3. X-ray Diffraction
3. Experimental Section
3.1. Expression and Purification of Wild-Type and Variant Forms of rHvExoI
3.2. Enzyme Assays
3.3. Crystallisation of Wild-Type and Variant Forms of rHvExoI
3.4. X-ray Data Collection and Processing
4. Conclusions
Acknowledgments
References
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Wild-type | Glu220Ala | Trp434Ala | Arg158Ala/Glu161Ala | |
---|---|---|---|---|
Unique reflections | 119968 | 74171 | 64460 | 107601 |
Resolution a (Å) | 1.57 (1.6−1.57) | 1.90 (1.94−1.90) | 1.95 (1.98−1.95) | 1.65 (1.68−1.65) |
Mean multiplicity a,b | 29 (26) | 27 (16) | 27 (16) | 26 (12) |
Completeness a,b (%) | 99 (86) | 99 (88) | 100 (100) | 99.8 (98) |
Mean <I/σ/(I)> a | 71.2 (5.6) | 54.8 (4.3) | 37.0 (2.2) | 58.7 (2.3) |
Rmerge a–c (%) | 6.7 (47) | 8.9 (64) | 10.1 (87) | 5.6 (82) |
a = b (Å) d | 99.2 | 100.2 | 100.1 | 100.8 |
c (Å) d | 183.5 | 183.2 | 183.6 | 183.2 |
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Luang, S.; Cairns, J.R.K.; Streltsov, V.A.; Hrmova, M. Crystallisation of Wild-Type and Variant Forms of a Recombinant Plant Enzyme β-D-Glucan Glucohydrolase from Barley (Hordeum vulgare L.) and Preliminary X-ray Analysis. Int. J. Mol. Sci. 2010, 11, 2759-2769. https://doi.org/10.3390/ijms11072759
Luang S, Cairns JRK, Streltsov VA, Hrmova M. Crystallisation of Wild-Type and Variant Forms of a Recombinant Plant Enzyme β-D-Glucan Glucohydrolase from Barley (Hordeum vulgare L.) and Preliminary X-ray Analysis. International Journal of Molecular Sciences. 2010; 11(7):2759-2769. https://doi.org/10.3390/ijms11072759
Chicago/Turabian StyleLuang, Sukanya, James R. Ketudat Cairns, Victor A. Streltsov, and Maria Hrmova. 2010. "Crystallisation of Wild-Type and Variant Forms of a Recombinant Plant Enzyme β-D-Glucan Glucohydrolase from Barley (Hordeum vulgare L.) and Preliminary X-ray Analysis" International Journal of Molecular Sciences 11, no. 7: 2759-2769. https://doi.org/10.3390/ijms11072759