Direct demonstration of NCAM cis-dimerization and inhibitory effect of palmitoylation using the BRET technique |
| |
| Authors: | Nikolaj Kulahin Lars Groth Grunnet Morten Lundh Dan Ploug Christensen Rasmus Jorgensen Anders Heding Nils Billestrup Vladimir Berezin Elisabeth Bock Thomas Mandrup-Poulsen |
| |
| Institution: | aProtein Laboratory, Department of Neuroscience and Pharmacology, Institute of Health Sciences, University of Copenhagen, Denmark;bENKAM Pharmaceuticals A/S, Copenhagen, Denmark;cReceptor Systems Biology Laboratory, Hagedorn Institute, Gentofte, Denmark;dDepartment of Translational Diabetology, Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark;eCore Unit for Medical Research Methodology, Department of Biomedical Sciences, University of Copenhagen, Denmark;f7TM Pharma, Hørsholm, Denmark;gGLP-1 & Obesity, Novo Nordisk A/S, Måløv, Denmark;hProtein Chemistry Biopharm, Novo Nordisk A/S, Måløv, Denmark |
| |
| Abstract: | 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 (BRET2) technique to directly demonstrate that full-length NCAM cis-homodimerizes in living cells. Based on BRET250 values we suggest that the intra-cellular part of NCAM inhibits cis-dimerization, an effect mainly dependent on the palmitoylation sites.Structured summaryMINT-8071463: NCAM140 (uniprotkb:P13591-1) physically interacts (MI:0915) with NCAM140 (uniprotkb:P13591-1) by bioluminescence resonance energy transfer (MI:0012)MINT-8071506: NCAM180 (genbank_protein_gi:119587605) physically interacts (MI:0915) with NCAM-180 (genbank_protein_gi:119587605) by bioluminescence resonance energy transfer (MI:0012)MINT-8071483: NCAM140 (uniprotkb:P13591-1) physically interacts (MI:0915) with NCAM140 (uniprotkb:P13591-1) by competition binding (MI:0405) |
| |
| Keywords: | Neural cell adhesion molecule Homophilic binding cis-interaction Bioluminescence resonance energy transfer Palmitoylation |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
