Characterization of the interaction of IpaB and IpaD, proteins required for entry of Shigella flexneri into epithelial cells, with a lipid membrane

Citation
C. De Geyter et al., Characterization of the interaction of IpaB and IpaD, proteins required for entry of Shigella flexneri into epithelial cells, with a lipid membrane, EUR J BIOCH, 267(18), 2000, pp. 5769-5776
Citations number
34
Categorie Soggetti
Biochemistry & Biophysics
Journal title
EUROPEAN JOURNAL OF BIOCHEMISTRY
ISSN journal
00142956 → ACNP
Volume
267
Issue
18
Year of publication
2000
Pages
5769 - 5776
Database
ISI
SICI code
0014-2956(200009)267:18<5769:COTIOI>2.0.ZU;2-F
Abstract
Entry of Shigella flexneri into epithelial cells and lysis of the phagosome involve the IpaB, IpaC, acid IpaD proteins, which are secreted by type III secretion machinery. We report here the purification of IpaB and IpaD and the characterization of their lipid-binding properties as a function of pH. The interaction of IpaB with the membrane was quite independent of the pH whereas that of IpaD took place only at low pH. To support the data obtaine d with the purified proteins, we designed a system in which protein secreti on by live bacteria was induced in the presence of liposomes, thereby allow ing interaction of proteins with lipids directly after secretion and bypass ing any purification step. In these conditions, both IpaB and IpaC, as well as minor amounts of IpaA and IpgD, were associated with the membrane and t he ratio of IpaB to IpaC was modulated by the pH. The relevance of these re sults with respect to the dual roles of IpaB, IpaC and IpaD in induction of membrane ruffles and lysis of the endosomal membrane is discussed.