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Direct demonstration of NCAM cis-dimerization and inhibitory effect of palmitoylation using the BRET2 technique.

Kulahin N., Grunnet L.G., Lundh M., Christensen D.P., Jorgensen R., Heding A., Billestrup N., Berezin V., Bock E., Mandrup-Poulsen T.

Biological activity of the neural cell adhesion molecule (NCAM) depends on both adhesion and activation of intra-cellular signaling. Based on in vitro experiments with truncated extra-cellular domains, several models describing homophilic NCAM trans- and cis-interactions have been proposed. However, cis-dimerization in living cells has not been shown directly and the role of the cytoplasmic part in NCAM dimerization is poorly understood. Here, we used the bioluminescence resonance energy transfer (BRET(2)) technique to directly demonstrate that full-length NCAM cis-homodimerizes in living cells. Based on BRET(2)50 values we suggest that the intra-cellular part of NCAM inhibits cis-dimerization, an effect mainly dependent on the palmitoylation sites.

FEBS Lett. 585:58-64(2011) [PubMed] [Europe PMC]

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