Proteins can be phosphorylated at neighboring sites resulting in different functional states, and studying the regulation of these sites has been challenging. Here we present Thesaurus, a search engine that detects and quantifies phosphopeptide positional isomers from parallel reaction monitoring and data-independent acquisition mass spectrometry experiments. We apply Thesaurus to analyze phosphorylation events in the PI3K/AKT signaling pathway and show neighboring sites with distinct regulation.
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http://dx.doi.org/10.1038/s41592-019-0498-4 | DOI Listing |
Eur Biophys J
November 2024
Department of Internal Medicine, Suzhou Wujiang District Children Hospital, Soochow University, Suzhou, 215200, China.
Human Snk is an evolutionarily conserved serine/threonine kinase essential for the maintenance of endocrine stability. The protein consists of a N-terminal catalytic domain and a C-terminal polo-box domain (PBD) that determines subcellular localization and substrate specificity. Here, an integrated strategy is described to explore the vast structural diversity space of Snk PBD-binding phosphopeptides at a molecular level using machine learning modeling, annealing optimization, dynamics simulation, and energetics rescoring, focusing on the recognition specificity and motif preference of the Snk PBD domain.
View Article and Find Full Text PDFJ Chromatogr A
January 2025
College of Chemistry, Jilin University, Changchun 130012, China. Electronic address:
Protein phosphorylation is a highly prevalent post-translational modification that holds a vital position in numerous physiological processes. Prior to mass spectrometry detection, the enrichment of phosphopeptides is critically significant due to their susceptibility to interference from abundant non-phosphopeptides. In this study, the magnetic nanocomposite (FeO@NE@PL) was successfully synthesized and characterized.
View Article and Find Full Text PDFJ Am Chem Soc
May 2024
Institut für Chemie, Humboldt-Universität zu Berlin, Brook-Taylor-Straße 2, 12489 Berlin, Germany.
Phosphorylation is a major constituent of the CTD code, which describes the set of post-translational modifications on 52 repeats of a YSPTSPS consensus heptad that orchestrates the binding of regulatory proteins to the C-terminal domain (CTD) of RNA polymerase II. Phospho-specific antibodies are used to detect CTD phosphorylation patterns. However, their recognition repertoire is underexplored due to limitations in the synthesis of long multiphosphorylated peptides.
View Article and Find Full Text PDFACS Omega
January 2024
Department of Biomedical Sciences, Creighton University, Omaha, Nebraska 68178, United States.
14-3-3ε is involved in various types of malignancies by increasing cell proliferation, promoting cell invasion, or inhibiting apoptosis. In cutaneous squamous cell carcinoma (cSCC), 14-3-3ε is overexpressed and mislocalized from the nucleus to the cytoplasm where it interacts with the cell division cycle 25 A (CDC25A) and suppresses apoptosis. Hence, inhibition of the 14-3-3ε-CDC25A interaction is an attractive target for promoting apoptosis in cSCC.
View Article and Find Full Text PDFNat Commun
December 2023
Department of Chemical and Structural Biology, The Weizmann Institute of Science, Rehovot, 7610001, Israel.
Peptide-based covalent probes can target shallow protein surfaces not typically addressable using small molecules, yet there is a need for versatile approaches to convert native peptide sequences into covalent binders that can target a broad range of residues. Here we report protein-based thio-methacrylate esters-electrophiles that can be installed easily on unprotected peptides and proteins via cysteine side chains, and react efficiently and selectively with cysteine and lysine side chains on the target. Methacrylate phosphopeptides derived from 14-3-3-binding proteins irreversibly label 14-3-3σ via either lysine or cysteine residues, depending on the position of the electrophile.
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