ArticleProceedings of the National Academy of Sciences of the United States of America2001
Galpha i3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 49 citations in OpenAlex.
- Heterotrimeric G protein signaling without GPCRs: The Gα-binding-and-activating (GBA) motif.The Journal of biological chemistry · 2024Review
- A biochemical and genetic discovery pipeline identifies PLCδ4b as a nonreceptor activator of heterotrimeric G-proteins.The Journal of biological chemistry · 2018Article
- Specific inhibition of GPCR-independent G protein signaling by a rationally engineered protein.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Rapid kinetic BRET measurements to monitor G protein activation by GPCR and non-GPCR proteins.Methods in cell biology · 2017Article
- Activation of Gαi at the Golgi by GIV/Girdin imposes finiteness in Arf1 signaling.Developmental cell · 2015Article
- Multimodular biosensors reveal a novel platform for activation of G proteins by growth factor receptors.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Gαs regulates the post-endocytic sorting of G protein-coupled receptors.Nature communications · 2014Article
- Regulation of the G-protein regulatory-Gαi signaling complex by nonreceptor guanine nucleotide exchange factors.The Journal of biological chemistry · 2013Article
- Serine protease activity of calnuc: regulation by Zn2+ and G proteins.The Journal of biological chemistry · 2013Article
- Non-canonical signaling and localizations of heterotrimeric G proteins.Cellular signalling · 2012Review
- G Protein binding sites on Calnuc (nucleobindin 1) and NUCB2 (nucleobindin 2) define a new class of G(alpha)i-regulatory motifs.The Journal of biological chemistry · 2011Article
- Regulation of the AGS3·G{alpha}i signaling complex by a seven-transmembrane span receptor.The Journal of biological chemistry · 2010Article
- Calnuc plays a role in dynamic distribution of Galphai but not Gbeta subunits and modulates ACTH secretion in AtT-20 neuroendocrine secretory cells.Molecular neurodegeneration · 2009Article
- The PDZ and band 4.1 containing protein Frmpd1 regulates the subcellular location of activator of G-protein signaling 3 and its interaction with G-proteins.The Journal of biological chemistry · 2008Article
- Activation of Galphai3 triggers cell migration via regulation of GIV.The Journal of cell biology · 2008Article
- Promotion of G alpha i3 subunit down-regulation by GIPN, a putative E3 ubiquitin ligase that interacts with RGS-GAIP.Proceedings of the National Academy of Sciences of the United States of America · 2003Article
- In situ analysis of spatial relationships between proteins of the nuclear pore complex.Biophysical journal · 2002Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
Galphai3 is found both on the plasma membrane and on Golgi membranes. Calnuc, an EF hand protein, binds both Galphai3 and Ca(2+) and is found both in the Golgi lumen and in the cytoplasm. To investigate whether Galphai3 binds calnuc in living cells and where this interaction takes place we performed fluorescence resonance energy transfer (FRET) analysis between Galphai3 and calnuc in COS-7 cells expressing Galphai3-yellow fluorescent protein (YFP) and calnuc-cyan fluorescent protein (CFP). The tagged proteins have the same localization as the endogenous, nontagged proteins. When Galphai3-YFP and calnuc-CFP are coexpressed, a FRET signal is detected in the Golgi region, but no FRET signal is detected on the plasma membrane. FRET is also seen within the Golgi region when Galphai3 is coexpressed with cytosolic calnuc(DeltaN2-25)-CFP lacking its signal sequence. No FRET signal is detected when Galphai3(DeltaC12)-YFP lacking the calnuc-binding region is coexpressed with calnuc-CFP or when Galphai3-YFP and calnuc(DeltaEF-1,2)-CFP, which is unable to bind Galphai3, are coexpressed. Galphai3(G2AC3A)-YFP lacking its lipid anchors is localized in the cytoplasm, and no FRET signal is detected when it is coexpressed with wild-type calnuc-CFP. These results indicate that cytosolic calnuc binds to Galphai3 on Golgi membranes in living cells and that Galphai3 must be anchored to the cytosolic surface of Golgi membranes via lipid anchors for the interaction to occur. Calnuc has the properties of a Ca(2+) sensor protein capable of binding to and potentially regulating interactions of Galphai3 on Golgi membranes.
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