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
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.