| Annotated protein: | Protein numb homolog. Gene symbol: NUMB. Taxonomy: Rattus norvegicus (Rat). Uniprot ID: Q2LC84 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ NUMB |
| Ontology domain: | Cellular Component |
| SynGO term: | postsynaptic density (GO:0014069) |
| Synapse type(s): | hippocampus, glutamatergic |
| Annotated paper: | Nishimura T, et al. "Role of numb in dendritic spine development with a Cdc42 GEF intersectin and EphB2" Mol Biol Cell. 2006 Mar;17(3):1273-85 PMID:16394100 |
| Figure(s): | Fig.1, 6 |
| Annotation description: | Fig.1: "Localization of Numb in dendritic spines" - " At high magnification, Numb were stained at puncta along with dendrites of layer V pyramidal neurons in the cerebral cortex and of the pyramidal neurons in the CA1 region of hippocampus (Figure 1D). These Numb puncta were largely overlapped with the postsynaptic marker PSD95, but not with the presynaptic marker synaptophysin. - "Although small puncta were distributed throughout the cell body, axon, and dendrites, large puncta were specifically localized at the dendritic spines (Figure 1G). The presence of Numb at postsynaptic sites was confirmed by its colocalization with F-actin, which accumulates in spines, and its overlap localization with PSD-95, a major component of the PSD, but appeared not with synaptophysin (see Figure 4B). Further complementary biochemical studies confirmed that a proportion of endogenous Numb is localized at the postsynapse. We isolated SPM from rat cerebral cortices and subsequently prepared PSD fractions. Numb concentrated in SPM fractions with PSD-95 and NMDA-type glutamate receptor NR2B (see Figure 6A). Approximately 50% of Numb evidently existed in the insoluble PSD pellets even after two rounds of 0.5% Triton extraction. Further extraction of Triton-insoluble complexes with 3% N-lauroyl sarcosine solubilized ~30% of the Numb present, leaving 70% in the PSD core. As expected, PSD-95 is concentrated in all insoluble, detergent-extracted PSD pellets, whereas synaptophysin is totally solubilized upon treatment of SPM with Triton." Fig.6: "Numb associates with EphB2 and NMDA receptors at dendritic spines" - "Both NMDA-type glutamate receptors NR1 and NR2B were efficiently precipitated with GST-Numb-full, GST-Numb-PTB, GST-Numb-PRR, and GST-intersectin-SH3 even in the presence of ionic detergents (Figure 6B)." - "Interestingly, GST-Numb fragments did not precipitate PSD95 in the presence of NMDA receptors, whereas GST-intersectin-SH3 did. To examine the physiological association of Numb with these receptors, we prepared isolated SPM lysate, which was solubilized with ionic-detergent, and precipitated Numb-interacting receptors by anti-Numb antibody. Numb immunoprecipitated NR1, NR2B, and EphB2 from the solubilized SPM fraction (Figure 6C). " |
| Evidence tracking, Biological System: | Intact tissue Cultured neurons Cell-free system |
| Evidence tracking, Protein Targeting: | Over-expression Antibody (detection) |
| Evidence tracking, Experiment Assay: | Confocal IP + WB/MSMS |
| Annotator(s): | Pim van Nierop (ORCID:0000-0003-0593-3443) Guus Smit (ORCID:0000-0002-2286-1587) Matthijs Verhage (ORCID:0000-0002-2514-0216) |
| Lab: | Department of Functional Genomics, Department of Molecular and Cellular Neurobiology, Center for Neurogenomics and Cognitive Research, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands |
| SynGO annotation ID: | 2863 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |