Article ; Online: Protocol to identify human subcellular alternative protein interactions using cross-linking mass spectrometry.
STAR protocols
2023 Volume 4, Issue 3, Page(s) 102380
Abstract: Since the start of mass-spectrometry-based proteomics, proteins from non-referenced open reading frames or alternative proteins (AltProts) have been overlooked. Here, we present a protocol to identify human subcellular AltProt and decipher some ... ...
Abstract | Since the start of mass-spectrometry-based proteomics, proteins from non-referenced open reading frames or alternative proteins (AltProts) have been overlooked. Here, we present a protocol to identify human subcellular AltProt and decipher some interactions using cross-linking mass spectrometry. We describe steps for cell culture, in cellulo cross-link, subcellular extraction, and sequential digestion. We then detail both liquid chromatography-tandem mass spectrometry and cross-link data analyses. The implementation of a single workflow allows the non-targeted identification of signaling pathways involving AltProts. For complete details on the use and execution of this protocol, please refer to Garcia-del Rio et al. |
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MeSH term(s) | Humans ; Proteins/metabolism ; Mass Spectrometry ; Proteomics/methods ; Signal Transduction |
Chemical Substances | Proteins |
Language | English |
Publishing date | 2023-06-28 |
Publishing country | United States |
Document type | Journal Article |
ISSN | 2666-1667 |
ISSN (online) | 2666-1667 |
DOI | 10.1016/j.xpro.2023.102380 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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