Synapse associated protein 102 is a novel binding partner to the cytoplasmic terminus of neurone-glial related cell adhesion molecule.
Neurone glial-related cell adhesion molecule (NrCAM) is a member of the L1 family of transmembrane cell adhesion receptors which are involved in the development and function of the mammalian nervous system. How these receptors interact with intracellular signalling pathways is not understood. To date the only identified binding partner to the cytoplasmic terminus of NrCAM is ankyrin G. We screened a developing rat brain cDNA yeast two-hybrid library with the cytoplasmic domain of NrCAM to identify further intracellular binding partners. We identified synapse associated protein 102 (SAP102) as a new binding partner for NrCAM. The interaction was confirmed biochemically using glutathione S-transferase (GST)-pull-down and tandem affinity purification, and also immunocytochemically as NrCAM and SAP102 co-localized in COS-7 and cerebellar granule cells. Binding was specific to NrCAM as neither neurofascin nor L1 bound SAP102, and this interaction was reliant on the last three amino acids of NrCAM. Additionally, NrCAM constructs whose last three amino acids had been deleted appeared to have a dominant negative effect on neurite extension of cerebellar granule cells. This is the first interaction reported for NrCAM, and its association with SAP102 suggests that it is part of a larger complex which can interact with many different signalling pathways.
Bute Medical Building, School of Biology, University of St Andrews, UK.
Fleur
F
Davey
Maria
M
Hill
Julien
J
Falk
Nathalie
N
Sans
Frank J
FJ
Gunn-Moore
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D019062:Cell Adhesion Molecules, Neuron-Glia / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D016922:Cellular Senescence; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D002522:Chlorocebus aethiops; D005982:Glutathione Transferase / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D051381:Rats; D014162:Transfection; D020798:Two-Hybrid System Techniques; D015003:Yeasts
D019062:Cell Adhesion Molecules, Neuron-Glia; C101195:Dlg3 protein, rat; D009479:Neuropeptides; D010446:Peptide Fragments; D005982:Glutathione Transferase