Novel strategy to study arginine deiminase partners in the parasite Giardia lamblia

TOUZ MC; ZLOCOWSKI, N; MAYOL, G; TOUZ MC; ROPOLO AS

Congreso; saib; 2014

Giardia lamblia is a ubiquitous unicellular parasite of humans and other vertebrates that commonly causes diarrhea and gastrointestinal upset. During the last years, we have been studying the multiple roles played by the enzyme arginine deiminase (ADI) during growth and differentiation of this parasite. An interesting feature is the capacity of ADI to interact with different partners from surface protein to histones depending on its localization during the cell cycle of Giardia. In this study, we develop a novel strategy to further analyzed ADI partners, by generating monoclonal antibodies (mAb) against in vivo immunoprecipitated Giardia ADI. By using ADI-HA as immunogen, obtained from immunoprecipitation using anti-HA mAb, we obtained not only specific mAbs against ADI but also to other proteins that co-immunoprecipitated with it. It is important to highlight the reproducibility of this method using different protein baits, which, in combination with MS-MS, allowed to obtain specific mAbs and also to disclose the architecture of particular protein networks.