Metabolic labeling of mammalian organisms with stable isotopes for quantitative proteomic analysis.
To quantify proteins on a global level from mammalian tissue, a method was developed to metabolically introduce 15N stable isotopes into the proteins of Rattus norvegicus for use as internal standards. The long-term metabolic labeling of rats with a diet enriched in 15N did not result in adverse health consequences. The average 15N amino acid enrichments reflected the relative turnover rates in the different tissues and ranged from 74.3 mpe in brain to 92.2 mpe in plasma. Using the 15N-enriched liver as a quantitative internal standard, changes in individual protein levels in response to cycloheximide treatment were measured for 310 proteins. These measurements revealed 127 proteins with altered protein level (p < 0.05). Most proteins with altered level have previously reported functions involving xenobiotic metabolism and protein-folding machinery of the endoplasmic reticulum. This approach is a powerful tool for the global quantitation of proteins, is capable of measuring proteome-wide changes in response to a drug, and will be useful for studying animal models of disease.
Wu
Christine C
CC
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
MacCoss
Michael J
MJ
Howell
Kathryn E
KE
Matthews
Dwight E
DE
Yates
John R
JR
D000818:Animals; D009587:Nitrogen Isotopes; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D051381:Rats
D009587:Nitrogen Isotopes; D011506:Proteins; D020543:Proteome