Cargando…
A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface
We demonstrate a two-hybrid assay based on antibody micropatterns to study protein-protein interactions at the cell surface of major histocompatibility complex class I (MHC I) proteins. Anti-tag and conformation-specific antibodies are used for individual capture of specific forms of MHC I proteins...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6125123/ https://www.ncbi.nlm.nih.gov/pubmed/30180933 http://dx.doi.org/10.7554/eLife.34150 |
_version_ | 1783353115069644800 |
---|---|
author | Dirscherl, Cindy Hein, Zeynep Ramnarayan, Venkat Raman Jacob-Dolan, Catherine Springer, Sebastian |
author_facet | Dirscherl, Cindy Hein, Zeynep Ramnarayan, Venkat Raman Jacob-Dolan, Catherine Springer, Sebastian |
author_sort | Dirscherl, Cindy |
collection | PubMed |
description | We demonstrate a two-hybrid assay based on antibody micropatterns to study protein-protein interactions at the cell surface of major histocompatibility complex class I (MHC I) proteins. Anti-tag and conformation-specific antibodies are used for individual capture of specific forms of MHC I proteins that allow for location- and conformation-specific analysis by fluorescence microscopy. The assay is used to study the in cis interactions of MHC I proteins at the cell surface under controlled conditions and to define the involved protein conformations. Our results show that homotypic in cis interactions occur exclusively between MHC I free heavy chains, and we identify the dissociation of the light chain from the MHC I protein complex as a condition for MHC I in cis interactions. The functional role of these MHC I protein-protein interactions at the cell surface needs further investigation. We propose future technical developments of our two-hybrid assay for further analysis of MHC I protein-protein interactions. |
format | Online Article Text |
id | pubmed-6125123 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-61251232018-09-07 A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface Dirscherl, Cindy Hein, Zeynep Ramnarayan, Venkat Raman Jacob-Dolan, Catherine Springer, Sebastian eLife Cell Biology We demonstrate a two-hybrid assay based on antibody micropatterns to study protein-protein interactions at the cell surface of major histocompatibility complex class I (MHC I) proteins. Anti-tag and conformation-specific antibodies are used for individual capture of specific forms of MHC I proteins that allow for location- and conformation-specific analysis by fluorescence microscopy. The assay is used to study the in cis interactions of MHC I proteins at the cell surface under controlled conditions and to define the involved protein conformations. Our results show that homotypic in cis interactions occur exclusively between MHC I free heavy chains, and we identify the dissociation of the light chain from the MHC I protein complex as a condition for MHC I in cis interactions. The functional role of these MHC I protein-protein interactions at the cell surface needs further investigation. We propose future technical developments of our two-hybrid assay for further analysis of MHC I protein-protein interactions. eLife Sciences Publications, Ltd 2018-09-05 /pmc/articles/PMC6125123/ /pubmed/30180933 http://dx.doi.org/10.7554/eLife.34150 Text en © 2018, Dirscherl et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Cell Biology Dirscherl, Cindy Hein, Zeynep Ramnarayan, Venkat Raman Jacob-Dolan, Catherine Springer, Sebastian A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface |
title | A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface |
title_full | A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface |
title_fullStr | A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface |
title_full_unstemmed | A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface |
title_short | A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface |
title_sort | two-hybrid antibody micropattern assay reveals specific in cis interactions of mhc i heavy chains at the cell surface |
topic | Cell Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6125123/ https://www.ncbi.nlm.nih.gov/pubmed/30180933 http://dx.doi.org/10.7554/eLife.34150 |
work_keys_str_mv | AT dirscherlcindy atwohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT heinzeynep atwohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT ramnarayanvenkatraman atwohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT jacobdolancatherine atwohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT springersebastian atwohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT dirscherlcindy twohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT heinzeynep twohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT ramnarayanvenkatraman twohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT jacobdolancatherine twohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface AT springersebastian twohybridantibodymicropatternassayrevealsspecificincisinteractionsofmhciheavychainsatthecellsurface |