Cloning, expression, purification, crystallization and preliminary X-ray studies of the C-terminal domain of Rv3262 (FbiB) from Mycobacterium tuberculosis

• Main Authors: Mohamed Rehan, Aisyah, Paterson, Neil G., Baker, Edward N., Squire, Christopher J.
• Format: Article
• Language: English
• Published: Wiley-Blackwell Publishing, Inc. 2011
• Subjects: Q Science (General); QH301 Biology
• Online Access: http://irep.iium.edu.my/9893/4/CLONING%2CEXPRESSION%2C_PURIFICATION%2C_CRYSTALLIZATION_AND_PRELIMINARY_X-RAY_STUDIES_OF_THE_C-TERMINAL.pdf; http://irep.iium.edu.my/9893/; http://scripts.iucr.org/cgi-bin/paper?S1744309111028958

During cofactor F420 biosynthesis, the enzyme F420-[gamma]-glutamyl ligase (FbiB) catalyzes the addition of [gamma]-linked L-glutamate residues to form polyglutamylated F420 derivatives. In Mycobacterium tuberculosis, Rv3262 (FbiB) consists of two domains: an N-terminal domain from the F420 ligase superfamily and a C-terminal domain with sequence similarity to nitro-FMN reductase superfamily proteins. To characterize the role of the C-terminal domain of FbiB in polyglutamyl ligation, it has been purified and crystallized in an apo form. The crystals diffracted to 2.0 Å resolution using a synchrotron source and belonged to the tetragonal space group P41212 (or P43212), with unit-cell parameters a = b = 136.6, c = 101.7 Å, [alpha] = [beta] = [gamma] = 90°.