Analysis of Proteins Interacting with TRIP8b Adapter

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N. V. Popova^1*, A. N. Plotnikov², R. Kh. Ziganshin¹, I. E. Deyev¹, and A. G. Petrenko^1,2

¹Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117991 Moscow, Russia; fax: (495) 335-7103; E-mail: petrea01@med.nyu.edu; petrenko@ibch.ru; popova@ibch.ru

²New York University, 10016 New York, USA

^* To whom correspondence should be addressed.

Received November 12, 2007; Revision received February 13, 2008

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Calcium-independent receptor of latrotoxin (CIRL) is an orphan heptahelical receptor implicated in regulation of exocytosis. To characterize molecular mechanisms of CIRL functioning, we searched for its intracellular partners using the yeast two-hybrid SR system with the cytoplasmic C-terminal fragment of CIRL as bait. One of the interacting proteins was identified as TRIP8b, a putative cytosolic adapter protein with multiple tetratricopeptide repeats. To understand functional significance of CIRL-TRIP8b interaction, we further isolated TRIP8b-interacting proteins by affinity chromatography of brain extracts on immobilized recombinant TRIP8b. Sixteen proteins were identified by mass spectrometry in the purified preparations. Clathrin and subunits of AP2 complex appeared to be the major TRIP8b-interacting proteins. Our data suggest a role of TRIP8b in receptor-mediated endocytosis.

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KEY WORDS: CIRL/latrophilin, TRIP8b, endocytosis, clathrin

DOI: 10.1134/S0006297908060035

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