1. Crystallization and preliminary X-ray crystallographic analysis of a full-length active form of the Cry4Ba toxin from Bacillus thuringiensis.
- Author
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Thamwiriyasati, Niramon, Sakdee, Somsri, Chuankhayan, Phimonphan, Katzenmeier, Gerd, Chen, Chun-Jung, and Angsuthanasombat, Chanan
- Subjects
BACTERIAL protein crystallography ,BACILLUS thuringiensis ,X-ray crystallography ,CRYSTALLIZATION ,X-ray diffraction ,MUTANT proteins ,MOSQUITO larvae ,INSECTICIDES - Abstract
To obtain a complete structure of the Bacillus thuringiensis Cry4Ba mosquito-larvicidal protein, a 65 kDa functional form of the Cry4Ba-R203Q mutant toxin was generated for crystallization by eliminating the tryptic cleavage site at Arg203. The 65 kDa trypsin-resistant fragment was purified and crystallized using the sitting-drop vapour-diffusion method. The crystals belonged to the rhombohedral space group R32, with unit-cell parameters a = b = 184.62, c = 187.36 Å. Diffraction data were collected to at least 2.07 Å resolution using synchrotron radiation and gave a data set with an overall R
merge of 9.1% and a completeness of 99.9%. Preliminary analysis indicated that the asymmetric unit contained one molecule of the active full-length mutant, with a VM coefficient and solvent content of 4.33 Å3 Daโ1 and 71%, respectively. [ABSTRACT FROM AUTHOR]- Published
- 2010
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