| Annotated protein: | Rho guanine nucleotide exchange factor 2 (Guanine nucleotide exchange factor H1) (GEF-H1). Gene symbol: ARHGEF2. Taxonomy: Rattus norvegicus (Rat). Uniprot ID: Q5FVC2 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ ARHGEF2 |
| Ontology domain: | Cellular Component |
| SynGO term: | postsynaptic density, intracellular component (GO:0099092) |
| Synapse type(s): | cerebral cortex, glutamatergic hippocampus, glutamatergic |
| Annotated paper: | Kang MG, et al. "AMPA receptor and GEF-H1/Lfc complex regulates dendritic spine development through RhoA signaling cascade" Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3549-54 PMID:19208802 |
| Figure(s): | fig. 1C-D, S2, S3A, S4C-E |
| Annotation description: | Fig. 1C-D, S2. GEF-H1 coimmunoprecipitate with AMPA receptor subunit GluR1. Literal "In small-scale IPs of endogenous AMPA-R complexes using cortical neuronal cultures, GEF-H1 as well as stargazin (a known interactor of AMPA-Rs) was specifically coimmunoprecipitated with GluR1 as a part of AMPA-R complex in neurons. Neither GEF-H1 nor stargazin was detected in this co-IP when GluR1 antibody was blocked by the synthetic peptide (see Fig. 1C, Peptide +). For further analysis of GEF-H1 association with AMPA-R complex, GFP-tagged GEF-H1 (GFP-GEF-H1) was coexpressed with myctagged AMPA-R subunits in heterologous cells, and the GFPGEF-H1 was immunoprecipitated using an anti-GFP antibody (see Fig. 1D). Both GluR1 and GluR2 were specifically coimmunoprecipitated with GFP-GEF-H1. However, the efficiency of this co-IP with heterologous cell lysate was much lower than that with brain or neuronal lysates, suggesting that the interaction may be stabilized by other proteins or by posttranslational modifications in neurons. Surprisingly, a GluR1 mutant lacking the cytoplasmic C-terminal (R1Ctdeklta) was also coimmunoprecipitated with GFP-GEF-H1, indicating that the C-terminal of AMPA-R is not required for GEF-H1 binding (see Fig. 1D). On the other hand, GluR6 (a subunit of kainate receptors) was not coimmunoprecipitated with GFP-GEF-H1, indicating that the interaction between AMPA-Rs and GEF-H1 is specific (Fig. S2)." Fig. S3A, S4C-E. GEF-H1 is enriched in the PSD fractions. Triple immunofluorescence staining of cultured pyramidal hippocampal neurons using antibodies against GFP and GluR1 after transfection of GFP-GEF-H1 and mCherry demonstrated that GFP-GEF-H1 is localized in spines as puncta and the puncta are colocalized with GluR1 puncta in the spines. Together, these data suggest that GEF-H1 and AMPA-R may form a complex in the PSD. Literal: "Our biochemical and immunocytochemical analyses of GEF-H1 showed localization of GEF-H1 in postsynaptic density (PSD) and spines (Figs. S3A and S4C)." |
| Evidence tracking, Biological System: | Intact tissue Cultured neurons |
| Evidence tracking, Protein Targeting: | Over-expression Antibody (detection) |
| Evidence tracking, Experiment Assay: | Confocal Western blot Biochemical fractionation (generic) |
| Annotator(s): | Chiara Verpelli (ORCID:0000-0003-2949-9725) Carlo Sala (ORCID:0000-0003-0662-9523) |
| Lab: | CNR Neuroscience Institute Milan and Dept. of Biotechnology and Translational Medicine, University of Milan, 20129 Milan, Italy |
| SynGO annotation ID: | 1747 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |