title: cysTMTRAQ-An integrative method for unbiased thiol-based redox proteomics.
abstract: Protein redox regulation plays important roles in many biological processes. Protein cysteine thiols are sensitive to redox changes and may function as redox switches, which turn signaling and metabolic pathways on or off to ensure speedy responses to environmental stimuli or stresses. Here we report a novel integrative proteomics method called cysTMTRAQ that combines two types of isobaric tags, cysteine tandem mass tags and isobaric tag for relative and absolute quantification, in one experiment. The method not only enables simultaneous analysis of cysteine redox changes and total protein level changes, but also allows the determination of bona fide redox modified cysteines in proteins through the correction of protein turnover. Overlooking the factor of protein-level changes in the course of protein posttranslational modification experiments could lead to misleading results. The capability to analyze protein posttranslational modification dynamics and protein-level changes in one experiment will advance proteomic studies in many areas of biology and medicine.
authors.affiliation: From the ‡Department of Biology, Genetics Institute, University of Florida, Gainesville, Florida 32611; §Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida 32610;
authors.forename: Jennifer
authors.initials: J
authors.lastname: Parker
authors.affiliation: From the ‡Department of Biology, Genetics Institute, University of Florida, Gainesville, Florida 32611; §Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida 32610;
authors.forename: Kelly
authors.initials: K
authors.lastname: Balmant
authors.affiliation: From the ‡Department of Biology, Genetics Institute, University of Florida, Gainesville, Florida 32611;
authors.forename: Fanchao
authors.initials: F
authors.lastname: Zhu
authors.affiliation: From the ‡Department of Biology, Genetics Institute, University of Florida, Gainesville, Florida 32611;
authors.forename: Ning
authors.initials: N
authors.lastname: Zhu
authors.affiliation: From the ‡Department of Biology, Genetics Institute, University of Florida, Gainesville, Florida 32611; §Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida 32610; ‖Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida 32610 schen@ufl.edu.
authors.forename: Sixue
authors.initials: S
authors.lastname: Chen
mesh_terms: D001059:Apoproteins / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism*; D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000378:metabolism; D007768:Lactalbumin / Q000378:metabolism; D007782:Lactoglobulins / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001059:Apoproteins; D029968:Escherichia coli Proteins; D007782:Lactoglobulins; D014168:Transferrin; C028284:apotransferrin; D012710:Serum Albumin, Bovine; D007768:Lactalbumin; D009113:Muramidase; D001616:beta-Galactosidase; D003545:Cysteine