Identification of interaction partners for individual SH3 domains of Fas ligand associated members of the PCH protein family in T lymphocytes

Abstract

Pombe Cdc15 homology (PCH) family proteins are regarded as key elements for linking membrane-associated processes to cytoskeletal elements and thus play a major role in exo- and endocytosis and organelle trafficking. We previously reported that, via their SH3 domains, several members of the PCH proteins interact with the proline-rich region of Fas ligand (FasL, CD95L), a key death factor in immune cells. Since protein–protein interactions that govern the storage and transport of FasL-associated vesicles are largely unknown, the present study was performed to identify other potential binding partners for SH3 domains of FasL-interacting PCH proteins. To this end, individual SH3 domains were expressed as GST fusion proteins and used to precipitate associated proteins from leukemic T cell lines and activated human T cell blasts. 87 protein bands representing 34 individual proteins were identified by mass spectrometry. The presented list of candidate interactors not only highlights the role of PCH proteins as adapters between vesicular membranes and the cytoskeleton but also points to an involvement of these proteins in the regulation of signalling events in T lymphocytes.