Abstract
The abundance of cellular proteins is determined largely by the rate of transcription and translation coupled with the stability of individual proteins. Although we know a great deal about global transcript abundance, little is known about global protein stability. We present a highly parallel multiplexing strategy to monitor protein turnover on a global scale by coupling flow cytometry with microarray technology to track the stability of individual proteins within a complex mixture. We demonstrated the feasibility of this approach by measuring the stability of approximately 8000 human proteins and identifying proteasome substrates. The technology provides a general platform for proteome-scale analysis of protein turnover under various physiological and disease conditions.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Amino Acids / analysis
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Cell Cycle
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Cell Line
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DNA, Complementary
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Flow Cytometry
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Green Fluorescent Proteins / analysis
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Green Fluorescent Proteins / metabolism
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Half-Life
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Humans
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Luminescent Proteins / analysis
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Luminescent Proteins / metabolism
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Oligonucleotide Array Sequence Analysis
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Open Reading Frames
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Proteasome Endopeptidase Complex / metabolism*
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Protein Biosynthesis
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Protein Stability*
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Proteins / genetics
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Proteins / metabolism*
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Recombinant Fusion Proteins / metabolism
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Red Fluorescent Protein
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Transcription, Genetic
Substances
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Amino Acids
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DNA, Complementary
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Luminescent Proteins
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Proteins
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RNA, Messenger
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Recombinant Fusion Proteins
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enhanced green fluorescent protein
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Green Fluorescent Proteins
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Proteasome Endopeptidase Complex