title: Global ubiquitination analysis by SILAC in mammalian cells.
abstract: Ubiquitination is a versatile and dynamic posttranslational modification in cells, regulating almost all cellular events. With rapid developments of affinity capture reagents and high-resolution mass spectrometry, it is now feasible to globally analyze the ubiquitinated proteome (ubiquitome) using quantitative strategies, such as stable isotope labeling with amino acids in cell culture (SILAC). Here we describe in detail a SILAC protocol to profile the ubiquitome in mammalian cells including protein labeling, antibody-based enrichment, and analysis by mass spectrometry.
authors: Department of Structural Biology, St. Jude Proteomics Facility, St Jude Children's Research Hospital, Memphis, TN, 38105, USA.
authors: Zhiping
authors: Z
authors: Wu
authors: Chan Hyun
authors: CH
authors: Na
authors: Haiyan
authors: H
authors: Tan
authors: Junmin
authors: J
authors: Peng
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D006033:Glycylglycine / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D054875:Ubiquitination
chemical_list: D000596:Amino Acids; D010455:Peptides; D006033:Glycylglycine