Abstract

Article info 2017 5 (3)    005  (03)   pp.  118 ~ 121
Title Purification, crystallization and X-ray crystallographic analysis of the type VI secretion system accessory protein TagF from Pseudomonas aeruginosa
Authors Chang-Kyu Ok and Jeong Ho Chang*
Institutions Department of Biology Education, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 41566, South Korea. *Correspondence: jhcbio@knu.ac.kr
Abstract TagF is the product of the type VI secretion system (T6SS) accessory gene F, which is capable of regulating T6SS assembly in Pseudomonas aeruginosa. Experimental results suggest that TagF may post-translationally regulate the T6SS through phosphatase activity, but the physiological functions of TagF are not yet understood. To provide structural insight into TagF function and the regulatory mechanisms controlling the T6SS, N-terminally His6-tagged TagF was overexpressed, purified, and crystallized using hanging-drop vapor diffusion in a solution of 2.5 M NaCl and 0.1 M Bis-Tris propane (pH 7.0). X-ray diffraction data from TagF crystals was collected at a resolution of 2.7 Å. TagF crystals belonged to the space group P21212, with unit cell parameters a = 93.7, b = 92.4, c = 151.1 Å, and α = β = γ = 90°. TagF consists of four independent subunits in the asymmetric unit.