Abstract
Many proteins expressed on the cellular surface provide signaling and cell adhesion properties required for vital cellular functions. These binding interactions can occur between different but complementary proteins such as a ligand and receptor, or between the same protein acting as both ligand and receptor. The cell aggregation assay is a straightforward technique to identify homophilic interactions from such proteins. Here we describe the procedure for testing proteins via cell aggregation assays in HEK293T cells.
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References
Wojtowicz WM, Flanagan JJ, Millard SS et al (2004) Alternative splicing of Drosophila Dscam generates axon guidance receptors that exhibit isoform-specific homophilic binding. Cell 118:619–633. https://doi.org/10.1016/j.cell.2004.08.021
Brown MH (2002) Detection of low-affinity ligand-receptor interactions at the cell surface with fluorescent microspheres. Curr Protoc Immunol Chapter 18:Unit 18.2. https://doi.org/10.1002/0471142735.im1802s48
Brown MH, Preston S, Barclay AN (1995) A sensitive assay for detecting low-affinity interactions at the cell surface reveals no additional ligands for the adhesion pair rat CD2 and CD48. Eur J Immunol 25:3222–3228. https://doi.org/10.1002/eji.1830251204
Matthews BJ, Kim ME, Flanagan JJ et al (2007) Dendrite self-avoidance is controlled by Dscam. Cell 129:593–604. https://doi.org/10.1016/j.cell.2007.04.013
Yamagata M, Sanes JR (2008) Dscam and sidekick proteins direct lamina-specific synaptic connections in vertebrate retina. Nature 451:465–469. https://doi.org/10.1038/nature06469
Schreiner D, Weiner JA (2010) Combinatorial homophilic interaction between gamma-protocadherin multimers greatly expands the molecular diversity of cell adhesion. Proc Natl Acad Sci U S A 107:14893–14898. https://doi.org/10.1073/pnas.1004526107
Ito K, Nakamura H, Watanabe Y (2011) Protogenin mediates cell adhesion for ingression and re-epithelialization of paraxial mesodermal cells. Dev Biol 351:13–24. https://doi.org/10.1016/j.ydbio.2010.11.024
Visser JJ, Cheng Y, Perry SC et al (2015) An extracellular biochemical screen reveals that FLRTs and Unc5s mediate neuronal subtype recognition in the retina. elife 4:1–27. https://doi.org/10.7554/eLife.08149
Karadge UB, Gosto M, Nicotra ML (2015) Allorecognition proteins in an invertebrate exhibit homophilic interactions. Curr Biol 25:2845–2850. https://doi.org/10.1016/j.cub.2015.09.030
Katsamba P, Carroll K, Ahlsen G et al (2009) Linking molecular affinity and cellular specificity in cadherin-mediated adhesion. Proc Natl Acad Sci U S A 106:11594–11599. https://doi.org/10.1073/pnas.0905349106
Kasinrerk W, Tokrasinwit N, Phunpae P (1999) CD147 monoclonal antibodies induce homotypic cell aggregation of monocytic cell line U937 via LFA-1/ICAM-1 pathway. Immunology 96:184–192. https://doi.org/10.1046/j.1365-2567.1999.00653.x
Chappuis-Flament S, Wong E, Hicks LD et al (2001) Multiple cadherin extracellular repeats mediate homophilic binding and adhesion. J Cell Biol 154:231–243. https://doi.org/10.1083/jcb.200103143
Karadge UB, Gosto M, Nicotra ML et al (2015) Allorecognition proteins in an invertebrate exhibit homophilic interactions report allorecognition proteins in an invertebrate exhibit homophilic interactions. Curr Biol 25:2845–2850. https://doi.org/10.1016/j.cub.2015.09.030
Sano K, Tanihara H, Heimark RL et al (1993) Protocadherins: a large family of cadherin-related molecules in central nervous system. EMBO J 12:2249–2256
Obata S, Sago H, Mori N et al (1995) Protocadherin Pcdh2 shows properties similar to, but distinct from, those of classical cadherins. J Cell Sci 108(Pt 12):3765–3773
Reiss K, Maretzky T, Haas IG et al (2006) Regulated ADAM10-dependent ectodomain shedding of g-protocadherin C3 modulates cell-cell adhesion. J Biol Chem 281:21735–21744. https://doi.org/10.1074/jbc.M602663200
Frank M, Ebert M, Shan W et al (2005) Differential expression of individual gamma-protocadherins during mouse brain development. Mol Cell Neurosci 29:603–616. https://doi.org/10.1016/j.mcn.2005.05.001
Gallen B, Triana-Baltzer MB (2006) Retinoids, eye development, and maturation of visual function. J Neurobiol 66:677–686. https://doi.org/10.1002/neu
Morishita H, Umitsu M, Murata Y et al (2006) Structure of the cadherin-related neuronal receptor/protocadherin-α first extracellular cadherin domain reveals diversity across cadherin families. J Biol Chem 281:33650–33663. https://doi.org/10.1074/jbc.M603298200
Fernández-Monreal M, Kang S, GRP (2013) NIH public access. Magn Reson Imaging 31:477–479. https://doi.org/10.1016/j.immuni.2010.12.017.Two-stage
Thu CA, Chen WV, Rubinstein R et al (2014) Single-cell identity generated by combinatorial homophilic interactions between a, b,and g protocadherins. Cell 158:1045–1059. https://doi.org/10.1016/j.cell.2014.07.012
Rubinstein R, Thu CA, Goodman KM et al (2015) Molecular logic of neuronal self-recognition through protocadherin domain interactions. Cell 163:629–642. https://doi.org/10.1016/j.cell.2015.09.026
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Huene, A.L., Nicotra, M.L. (2022). Cell Aggregation Assays for Homophilic Interactions Between Cell Surface Proteins. In: Rast, J., Buckley, K. (eds) Immune Receptors. Methods in Molecular Biology, vol 2421. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1944-5_6
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DOI: https://doi.org/10.1007/978-1-0716-1944-5_6
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