Validation of the Applicability of In-Cell Fast Photochemical Oxidation of Proteins across Multiple Eukaryotic Cell Lines.

Journal of the American Society for Mass Spectrometry
Upneet KaurLisa M Jones

Abstract

Fast photochemical oxidation of proteins (FPOP), a hydroxyl radical-based protein footprinting method, coupled to mass spectrometry has been extensively used to study protein structure and protein-protein interactions in vitro. This method utilizes hydroxyl radicals to oxidatively modify solvent-accessible amino acids and has recently been demonstrated to modify proteins within live cells (IC-FPOP) and Caenorhabditis elegans. Here, we have expanded the application of IC-FPOP into a variety of commonly used cell lines to verify the applicability of the method across various cellular systems. IC-FPOP was able to successfully modify proteins in five different cell lines (Vero, HEK 293T, CHO, MCF-10A, and MCF-7). To increase the number of oxidatively modified proteins identified, we have also employed the use of offline high pH reversed-phase liquid chromatography (RPLC) followed by concatenation and online low-pH RPLC. The coupling of IC-FPOP to 2D-LC MS/MS resulted in a 1.7-fold increase in total identifications of oxidatively modified proteins, which expanded the dynamic range of the method. This work demonstrates the efficacy of using IC-FPOP to study protein-protein interactions in cells.

References

Jul 4, 2002·Annals of the New York Academy of Sciences·Ida Pucci-MinafraSalvatore Minafra
Jul 29, 2003·Journal of Mammary Gland Biology and Neoplasia·Hubert HondermarckJérôme Lemoine
Jan 13, 2005·Molecular & Cellular Proteomics : MCP·José M A MoreiraJulio E Celis
Nov 3, 2005·Journal of the American Society for Mass Spectrometry·David M Hambly, Michael L Gross
Mar 4, 2008·Seminars in Radiation Oncology·Dimitry S A Nuyten, Marc J van de Vijver
Mar 15, 2008·Cancer Research·Christopher G DanesDihua Yu
Jun 6, 2008·Molecular and Cellular Endocrinology·Dominique GalloGuy Leclercq
Sep 9, 2008·Applied Immunohistochemistry & Molecular Morphology : AIMM·Ying CaoConstance T Albarracin
May 29, 2009·Molecular & Cellular Proteomics : MCP·Yi ZhuSiu Kwan Sze
Dec 19, 2009·Nucleic Acids Research·Natalie A CooksonJeff Hasty
Sep 20, 2011·Trends in Biotechnology·Georgeta Crivat, Justin W Taraska
Sep 29, 2011·Seminars in Cell & Developmental Biology·Alexandra K Gardino, Michael B Yaffe
Oct 11, 2011·Seminars in Cell & Developmental Biology·Jing ZhaoHaian Fu
Feb 2, 2012·Expert Review of Proteomics·Yueyang ZhongBrandon T Ruotolo
Apr 3, 2012·Expert Review of Proteomics·Feng YangRichard D Smith
Oct 13, 2012·The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology·Ellen C Jensen
Oct 4, 2014·Science·Mikhail M SavitskiGerard Drewes
Jan 28, 2015·Proceedings of the National Academy of Sciences of the United States of America·William B MonteithGary J Pielak
Feb 14, 2015·Protein Science : a Publication of the Protein Society·Elisabetta Boeri Erba, Carlo Petosa
Feb 14, 2016·Analytical and Bioanalytical Chemistry·Aimee RinasLisa M Jones
Jun 17, 2016·Annual Review of Analytical Chemistry·Timothy K TobyNeil L Kelleher
Dec 3, 2016·Journal of the American Society for Mass Spectrometry·Aneika C Leney, Albert J R Heck
Mar 10, 2017·Molecular & Cellular Proteomics : MCP·Liwen Wang, Mark R Chance
Nov 21, 2018·Analytical Chemistry·Upneet KaurLisa M Jones
Apr 27, 2019·Analytical Chemistry·Jessica A Espino, Lisa M Jones
Jul 3, 2019·The Journal of Biological Chemistry·Danté T JohnsonLisa M Jones
Jan 15, 2020·Molecular Omics·Kellye A Cupp-Sutton, Si Wu

❮ Previous
Next ❯

Related Concepts

Related Feeds

Autoimmune Polyendocrine Syndromes

This feed focuses on a rare genetic condition called Autoimmune Polyendocrine Syndromes, which are characterized by autoantibodies against multiple endocrine organs. This can lead to Type I Diabetes.

Autoimmune Polyendocrinopathies

Autoimmune polyendocrinopathies, also called polyglandular autoimmune syndromes (PGASs), or polyendocrine autoimmune syndromes(PASs), are a heterogeneous group of rare diseases characterized by autoimmune activity against more than one endocrine organ, although non-endocrine organs can be affected. Discover the latest research on autoimmune polyendocrinopathies here.