Global protein stability profiling in mammalian cells.
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.
Yen
Hsueh-Chi Sherry
HC
Department of Genetics, Center for Genetics and Genomics, Brigham and Women's Hospital, Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115, USA.
Xu
Qikai
Q
Chou
Danny M
DM
Zhao
Zhenming
Z
Elledge
Stephen J
SJ
D000596:Amino Acids / Q000032:analysis; D002453:Cell Cycle; D002460:Cell Line; D018076:DNA, Complementary; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D014176:Protein Biosynthesis; D055550:Protein Stability; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014158:Transcription, Genetic; D000097573:Red Fluorescent Protein
D000596:Amino Acids; D018076:DNA, Complementary; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D046988:Proteasome Endopeptidase Complex; D011506:Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein