Abstract
Type III secretion (T3S) needle proteins are essential for the pathogenesis of many gram-negative bacteria. The needle component of the T3S system serves as the conduit for the translocation of effector proteins from the cytoplasm of many gram-negative bacteria into their target eukaryotic cells. Despite substantial advances that have been made in their characterization, a lot is still unknown about their interactions with other T3S system proteins and their roles in modulating host immune responses during infections. Critical to achieving this knowledge is the ability to isolate these needle proteins in their stable, native form. In this chapter, we describe a modified, streamlined isolation strategy for native forms of these T3S system needle proteins. We also present assays to detect the presence and quantification of these needle proteins.
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