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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...

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Detalles Bibliográficos
Autores principales: Dirscherl, Cindy, Hein, Zeynep, Ramnarayan, Venkat Raman, Jacob-Dolan, Catherine, Springer, Sebastian
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
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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.
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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
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