Premium
Crystallization and preliminary X‐ray analysis of the periplasmic domain of FliP, an integral membrane component of the bacterial flagellar type III protein‐export apparatus
Author(s) -
Fukumura Takuma,
Furukawa Yukio,
Kawaguchi Tatsuya,
SaijoHamano Yumiko,
Namba Keiichi,
Imada Katsumi,
Minamino Tohru
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14014678
Subject(s) - periplasmic space , crystallization , crystallography , thermotoga maritima , chemistry , membrane , homotetramer , transmembrane protein , signal peptide , biophysics , biochemistry , escherichia coli , biology , peptide sequence , gene , receptor , organic chemistry , protein subunit
The bacterial flagellar proteins are transported via a specific export apparatus to the distal end of the growing structure for their self‐assembly. FliP is an essential membrane component of the export apparatus. FliP has an N‐terminal signal peptide and is predicted to have four transmembrane (TM) helices and a periplasmic domain (FliP P ) between TM‐2 and TM‐3. In this study, FliP P from Thermotoga maritima (TmFliP P ) and its selenomethionine derivative (SeMet‐TmFliP P ) were purified and crystallized. TmFliP P formed a homotetramer in solution. Crystals of TmFliP P and SeMet‐TmFliP P were obtained by the hanging‐drop vapour‐diffusion technique with 2‐methyl‐2,4‐pentanediol as a precipitant. These two crystals grew in the hexagonal space group P 6 2 22 or P 6 4 22, with unit‐cell parameters a = b = 114.9, c = 193.8 Å. X‐ray diffraction data were collected from crystals of TmFliP P and SeMet‐TmFliP P to 2.4 and 2.8 Å resolution, respectively.