Open AccessCrystallization and preliminary X‐ray crystallographic analysis of a full‐length active form of the Cry4Ba toxin from Bacillus thuringiensis
Author(s) -
Thamwiriyasati Niramon,
Sakdee Somsri,
Chuankhayan Phimonphan,
Katzenmeier Gerd,
Chen ChunJung,
Angsuthanasombat Chanan
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110015344
Subject(s) - crystallization , crystallography , bacillus thuringiensis , chemistry , mutant , materials science , biology , biochemistry , gene , organic chemistry , bacteria , genetics
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 R 32, 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 V M coefficient and solvent content of 4.33 Å 3 Da −1 and 71%, respectively.