title: Recombinant Expression and Purification of the Shigella Translocator IpaB.
abstract: Type III secretion systems (T3SS) are highly conserved virulence factors employed by a large number of pathogenic gram-negative bacteria. Like many T3SS translocators, recombinant expression of the hydrophobic Shigella protein IpaB requires the presence of its cognate chaperone IpgC. Chaperone-bound IpaB is maintained in a nonfunctional state, which has hampered in vitro studies aimed at understanding molecular structure and function of this important class of T3SS proteins. Herein, we describe an expression and purification protocol that utilizes mild detergents to produce highly purified, homogeneous IpaB of defined oligomeric states.
authors.affiliation: Higuchi Biosciences Center, University of Kansas, Lawrence, KS, 66047, USA.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Barta
authors.affiliation: Kansas Department of Health and Environment Laboratories, Topeka, KS, 66620, USA.
authors.forename: Philip R
authors.initials: PR
authors.lastname: Adam
authors.affiliation: Department of Chemistry and Biochemistry, Utah State University, 300 Old Main Hill, Logan, UT, 84322, USA. nick.dickenson@usu.edu.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Dickenson
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins; D000069296:Type III Secretion Systems / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D000069296:Type III Secretion Systems; C087447:ipaB protein, Shigella; C094580:IpgC protein, Shigella
keywords: Chaperone; Detergent; IpaB; Membrane protein; Shigella; Translocator; Type III secretion system