Capture, Release, and Identification of Newly Synthesized Proteins for Improved Profiling of Functional Translatomes.

Biotin Azides Proteins / chemistry Peptides RNA, Messenger
O-propargyl-puromycin cleavable linker mTOR nascent proteins translation

Journal

Molecular & cellular proteomics : MCP
ISSN: 1535-9484
Titre abrégé: Mol Cell Proteomics
Pays: United States
ID NLM: 101125647

Informations de publication

Date de publication:
03 2023
Historique:
received: 27 07 2022
revised: 17 12 2022
accepted: 09 01 2023
medline: 28 3 2023
pubmed: 16 1 2023
entrez: 15 1 2023
Statut: ppublish

Résumé

New protein synthesis is regulated both at the level of mRNA transcription and translation. RNA-Seq is effective at measuring levels of mRNA expression, but techniques to monitor mRNA translation are much more limited. Previously, we reported results from O-propargyl-puromycin (OPP) labeling of proteins undergoing active translation in a 2-h time frame, followed by biotinylation using click chemistry, affinity purification, and on-bead digestion to identify nascent proteins by mass spectrometry (OPP-ID). As with any on-bead digestion protocol, the problem of nonspecific binders complicated the rigorous categorization of nascent proteins by OPP-ID. Here, we incorporate a chemically cleavable linker, Dde biotin-azide, into the protocol (OPP-ID

Identifiants

DOI: 10.1016/j.mcpro.2023.100497 PMID: 36642223 PMC: PMC9971285
pubmed: 36642223
pii: S1535-9476(23)00006-3
doi: 10.1016/j.mcpro.2023.100497
pmc: PMC9971285
pii:
doi:

Substances chimiques

Biotin 6SO6U10H04
Azides 0
Proteins 0
Peptides 0
RNA, Messenger 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

100497

Subventions

Organisme : Howard Hughes Medical Institute
Pays : United States

Informations de copyright

Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.

Auteurs

Nancy J Phillips (NJ)

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.

Bala M Vinaithirthan (BM)

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.

Juan A Oses-Prieto (JA)

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.

Robert J Chalkley (RJ)

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.

Alma L Burlingame (AL)

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA; Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California, USA. Electronic address: alb@cgl.ucsf.edu.

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Classifications MeSH

questionsmedicales.fr › Enzymes et coenzymes › Coenzymes › Biotine › Capture, Release, and Identification of Newly...
questionsmedicales.fr › Composés chimiques organiques › Azotures › Capture, Release, and Identification of Newly...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Capture, Release, and Identification of Newly...
questionsmedicales.fr › Acides aminés, peptides et protéines › Peptides › Capture, Release, and Identification of Newly...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN messager › Capture, Release, and Identification of Newly...