Open AccessPurification, crystallization and preliminary X‐ray analysis of FliT, a bacterial flagellar substrate‐specific export chaperone
Author(s) -
Kinoshita Miki,
Yamane Midori,
Matsunami Hideyuki,
Minamino Tohru,
Namba Keiichi,
Imada Katsumi
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109026736
Subject(s) - flagellum , chaperone (clinical) , crystallization , salmonella enterica , crystallography , biology , gene , regulator , chemistry , biophysics , escherichia coli , biochemistry , medicine , organic chemistry , pathology
The assembly process of the bacterial flagellum is coupled to flagellar gene expression. FliT acts not only as a flagellar type III substrate‐specific export chaperone for the filament‐capping protein FliD but also as a negative regulator that suppresses flagellar gene expression through its specific interaction with the master regulator FlhD 4 C 2 complex. In this study, FliT of Salmonella enterica serovar Typhimurium was expressed, purified and crystallized. Crystals of SeMet FliT were obtained by the sitting‐drop vapour‐diffusion technique with potassium/sodium tartrate as the precipitant. The crystals grew in the trigonal space group P 3 1 21 or P 3 2 21 and diffracted to 3.2 Å resolution. The anomalous difference Patterson map of the SeMet FliT crystal showed significant peaks in its Harker sections, indicating the usefulness of the derivative data for structure determination.