Asset Details
Do you wish to reserve the book?
Benchmarking common quantification strategies for large-scale phosphoproteomics
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?
Benchmarking common quantification strategies for large-scale phosphoproteomics
Do you wish to request the book?
Benchmarking common quantification strategies for large-scale phosphoproteomics
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
Benchmarking common quantification strategies for large-scale phosphoproteomics
2018
Overview
Comprehensive mass spectrometry (MS)-based proteomics is now feasible, but reproducible quantification remains challenging, especially for post-translational modifications such as phosphorylation. Here, we compare the most popular quantification techniques for global phosphoproteomics: label-free quantification (LFQ), stable isotope labeling by amino acids in cell culture (SILAC) and MS
2
- and MS
3
-measured tandem mass tags (TMT). In a mixed species comparison with fixed phosphopeptide ratios, we find LFQ and SILAC to be the most accurate techniques. MS
2
-based TMT yields the highest precision but lowest accuracy due to ratio compression, which MS
3
-based TMT can partly rescue. However, MS
2
-based TMT outperforms MS
3
-based TMT when analyzing phosphoproteome changes in the DNA damage response, since its higher precision and larger identification numbers allow detection of a greater number of significantly regulated phosphopeptides. Finally, we utilize the TMT multiplexing capabilities to develop an algorithm for determining phosphorylation site stoichiometry, showing that such applications benefit from the high accuracy of MS
3
-based TMT.
Quantitative phosphoproteomics has become a standard method in molecular and cell biology. Here, the authors compare performance and parameters of phosphoproteome quantification by LFQ, SILAC, and MS
2
-/MS
3
-based TMT and introduce a TMT-adapted algorithm for calculating phosphorylation site stoichiometry.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
