Biopolym. Cell. 2014; 30(1):37-41.
Structure and Function of Biopolymers
Interaction between adaptor proteins Ruk/CIN85 and Tks4 in normal and tumor cells of different tissue origins
1Bazalii A. V., 1Samoylenko A. A., 1Petukhov D. M., 2Rynditch A. V., 3Redowicz M.-J., 1Drobot L. B.
- Palladin Institute of Biochemistry, NAS of Ukraine
9, Leontovycha Str., Kyiv, Ukraine, 01601 - Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 - Nencki Institute of Experimental Biology, Polish Academy of Sciences
3, Pasteur Str., Warszawa, Poland, 02-093
Abstract
Aim. To study the interaction of adaptor protein Ruk/CIN85 SH3 domains with endogenous adaptor protein Tks4 in normal and tumor cells of different tissue origins. Methods. GST in vitro pull-down assay was performed using total cell lysates of cell lines of different origins. Results. Using GST in vitro pull-down assay, we have determined that SH3A domain of adaptor protein Ruk/CIN85 precipitated full-length form of adaptor protein Tks4 (Mr 120 kDa) from lysates of human breast (MCF-7, MDA-MB-231), melanoma (MM-4), colon (HT-29, DLD-1) tumor cells as well as from lysates of mouse Lewis lung carcinoma cells (LLC) and fibroblasts (NIH 3T3). It has been also revealed that all Ruk/CIN85 SH3 domains (A, B and C) with high efficiency precipitate the additional forms of Tks4 with Mr 75, 90 and 160 kDa from lysates of human colon carcinoma cells and mouse fibroblasts. The molecular nature of new multiple forms of Tks4 has not been determined to date. Conclusions. New multiple molecular forms of adaptor protein Tks4 with Mr 75, 90 and 160 kDa, able to interact with high affinity with SH3 domains of Ruk/CIN85, were identified using GST in vitro pull-down assay. The data obtained suggest that interaction between Ruk/CIN85 SH3 domains with Tks4 endogenous forms is determined by cellular context while a level of this interaction can be regulated in the course of physiological cellular responses.
Keywords: adaptor proteins, Ruk/CIN85, Tks4, SH3 domains, GST in vitro pull-down assay
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