title: A six-plex proteome quantification strategy reveals the dynamics of protein turnover.
abstract: MS1 full scan based quantification is one of the most popular approaches for large-scale proteome quantification. Typically only three different samples can be differentially labeled and quantified in a single experiment. Here we present a two stages stable isotope labeling strategy which allows six different protein samples (six-plex) to be reliably labeled and simultaneously quantified at MS1 level. Briefly in the first stage, isotope lysine-d0 (K0) and lysine-d4 (K4) are in vivo incorporated into different protein samples during cell culture. Then in the second stage, three of K0 and K4 labeled protein samples are digested by lysine C and in vitro labeled with light (2CH3), medium (2CD2H), and heavy (2(13)CD3) dimethyl groups, respectively. We demonstrated that this six-plex isotope labeling strategy could successfully investigate the dynamics of protein turnover in a high throughput manner.
authors: CAS Key Lab of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. wangfj@dicp.ac.cn
authors: Fangjun
authors: F
authors: Wang
authors: Kai
authors: K
authors: Cheng
authors: Xiaoluan
authors: X
authors: Wei
authors: Hongqiang
authors: H
authors: Qin
authors: Rui
authors: R
authors: Chen
authors: Jing
authors: J
authors: Liu
authors: Hanfa
authors: H
authors: Zou
mesh_terms: D002460:Cell Line; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D007553:Isotope Labeling; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012984:Software; D054875:Ubiquitination
chemical_list: D010455:Peptides; D011506:Proteins; D020543:Proteome
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