Asset Details
Do you wish to reserve the book?
Rapid and site-specific deep phosphoproteome profiling by data-independent acquisition without the need for spectral libraries
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Rapid and site-specific deep phosphoproteome profiling by data-independent acquisition without the need for spectral libraries
Do you wish to request the book?
Rapid and site-specific deep phosphoproteome profiling by data-independent acquisition without the need for spectral libraries
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Rapid and site-specific deep phosphoproteome profiling by data-independent acquisition without the need for spectral libraries
2020
Overview
Quantitative phosphoproteomics has transformed investigations of cell signaling, but it remains challenging to scale the technology for high-throughput analyses. Here we report a rapid and reproducible approach to analyze hundreds of phosphoproteomes using data-independent acquisition (DIA) with an accurate site localization score incorporated into Spectronaut. DIA-based phosphoproteomics achieves an order of magnitude broader dynamic range, higher reproducibility of identification, and improved sensitivity and accuracy of quantification compared to state-of-the-art data-dependent acquisition (DDA)-based phosphoproteomics. Notably, direct DIA without the need of spectral libraries performs close to analyses using project-specific libraries, quantifying > 20,000 phosphopeptides in 15 min single-shot LC-MS analysis per condition. Adaptation of a 3D multiple regression model-based algorithm enables global determination of phosphorylation site stoichiometry in DIA. Scalability of the DIA approach is demonstrated by systematically analyzing the effects of thirty kinase inhibitors in context of epidermal growth factor (EGF) signaling showing that specific protein kinases mediate EGF-dependent phospho-regulation.
Localizing phosphorylation sites by data-independent acquisition (DIA)-based proteomics is still challenging. Here, the authors develop algorithms for phosphosite localization and stoichiometry determination, and incorporate them into single-shot DIA-phosphoproteomics workflows.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 82/58
/ 96
/ 96/95
/ Chromatography, Liquid - methods
/ Computational Biology - methods
/ Epidermal Growth Factor - metabolism
/ High-Throughput Screening Assays - methods
/ Humanities and Social Sciences
/ Humans
/ Kinases
/ Phosphopeptides - metabolism
/ Protein Kinase Inhibitors - pharmacology
/ Protein Kinases - metabolism
/ Science
