Chemical crosslinkers enhance detection of receptor interactomes
Receptor function is dependent on interaction with various intracellular proteins that ensure the localization and signaling of the receptor. While a number of approaches have been optimized for the isolation, purification, and proteomic characterization of receptor-protein interaction networks (int...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2014-01-01
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| Series: | Frontiers in Pharmacology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fphar.2013.00171/full |
| _version_ | 1818149701174689792 |
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| author | Brian A Corgiat Jacob C Nordman Nadine eKabbani |
| author_facet | Brian A Corgiat Jacob C Nordman Nadine eKabbani |
| author_sort | Brian A Corgiat |
| collection | DOAJ |
| description | Receptor function is dependent on interaction with various intracellular proteins that ensure the localization and signaling of the receptor. While a number of approaches have been optimized for the isolation, purification, and proteomic characterization of receptor-protein interaction networks (interactomes) in cells, the capture of receptor interactomes and their dynamic properties remains a challenge. In particular, the study of interactome components that bind to the receptor with low affinity or can rapidly dissociate from the macromolecular complex is difficult. Here we describe how chemical crosslinking (CC) can aid in the isolation and proteomic analysis of receptor-protein interactions in cells. The addition of CC to standard affinity purification and mass spectrometry protocols boosts the power of protein capture within the proteomic assay and enables the identification of specific binding partners under various cellular and receptor states. The utility of CC in receptor interactome studies is highlighted for the nicotinic acetylcholine receptor (nAChR) as well as several other receptor types. A better understanding of receptors and their interactions with proteins spearheads molecular biology, informs an integral part of bench medicine which does help in drug development, drug action, and understanding the pathophysiology of disease. <br/><br/> |
| first_indexed | 2024-12-11T13:11:13Z |
| format | Article |
| id | doaj.art-e9abbf1020d64693bfe2fb6d6532f6ea |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-11T13:11:13Z |
| publishDate | 2014-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-e9abbf1020d64693bfe2fb6d6532f6ea2022-12-22T01:06:10ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122014-01-01410.3389/fphar.2013.0017172387Chemical crosslinkers enhance detection of receptor interactomesBrian A Corgiat0Jacob C Nordman1Nadine eKabbani2George Mason UniversityGeorge Mason UniversityGeorge Mason UniversityReceptor function is dependent on interaction with various intracellular proteins that ensure the localization and signaling of the receptor. While a number of approaches have been optimized for the isolation, purification, and proteomic characterization of receptor-protein interaction networks (interactomes) in cells, the capture of receptor interactomes and their dynamic properties remains a challenge. In particular, the study of interactome components that bind to the receptor with low affinity or can rapidly dissociate from the macromolecular complex is difficult. Here we describe how chemical crosslinking (CC) can aid in the isolation and proteomic analysis of receptor-protein interactions in cells. The addition of CC to standard affinity purification and mass spectrometry protocols boosts the power of protein capture within the proteomic assay and enables the identification of specific binding partners under various cellular and receptor states. The utility of CC in receptor interactome studies is highlighted for the nicotinic acetylcholine receptor (nAChR) as well as several other receptor types. A better understanding of receptors and their interactions with proteins spearheads molecular biology, informs an integral part of bench medicine which does help in drug development, drug action, and understanding the pathophysiology of disease. <br/><br/>http://journal.frontiersin.org/Journal/10.3389/fphar.2013.00171/fullMass Spectrometrynicotinic receptorsInteractomesignaling networkchemical crosslinking |
| spellingShingle | Brian A Corgiat Jacob C Nordman Nadine eKabbani Chemical crosslinkers enhance detection of receptor interactomes Frontiers in Pharmacology Mass Spectrometry nicotinic receptors Interactome signaling network chemical crosslinking |
| title | Chemical crosslinkers enhance detection of receptor interactomes |
| title_full | Chemical crosslinkers enhance detection of receptor interactomes |
| title_fullStr | Chemical crosslinkers enhance detection of receptor interactomes |
| title_full_unstemmed | Chemical crosslinkers enhance detection of receptor interactomes |
| title_short | Chemical crosslinkers enhance detection of receptor interactomes |
| title_sort | chemical crosslinkers enhance detection of receptor interactomes |
| topic | Mass Spectrometry nicotinic receptors Interactome signaling network chemical crosslinking |
| url | http://journal.frontiersin.org/Journal/10.3389/fphar.2013.00171/full |
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