Inverse labeling-mass spectrometry for the rapid identification of differentially expressed protein markers/targets.
Comparative proteomic studies can lead to the identification of protein markers for disease diagnostics and protein targets for potential disease interventions. An inverse labeling strategy based on the principle of protein stable isotope labeling and mass spectrometric detection has been successfully applied to three general protein labeling methods. In contrast to the conventional single experiment approach, two labeling experiments are performed in which the initial labeling is reversed in the second experiment. Signals from differentially expressed proteins will distinguish themselves by exhibiting a characteristic pattern of isotope intensity profile reversal that will lead to the rapid identification of these proteins. Application of the inverse labeling method is demonstrated using model systems for protein chemical labeling, protein proteolytic labeling, and protein metabolic labeling. The methodology has clear advantages which are illustrated in the various studies. The inverse labeling strategy permits quick focus on signals from differentially expressed proteins (markers/targets) and eliminates ambiguities caused by the dynamic range of detection. In addition, the inverse labeling approach enables the unambiguous detection of covalent changes of proteins responding to a perturbation.
Wang
Y Karen
YK
Central Technologies, Drug Discovery Research, Novartis Pharmaceuticals Corporation, 556 Morris Avenue, Summit, NJ 07901, USA. karen.wang@pharma.novartis.com
Quinn
Douglas F
DF
Ma
Zhixiang
Z
Fu
Emil W
EW
D002853:Chromatography, Liquid / Q000379:methods; D006868:Hydrolysis; D007553:Isotope Labeling; D013058:Mass Spectrometry / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
D011506:Proteins