Ross, P. L., Huang, Y. N., Marchese, J. N., Williamson, B., Parker, K., Hattan, S., Khainovski, N., Pillai, S., Dey, S., Daniels, S., Purkayastha, S., Juhasz, P., Martin, S., Bartlet-Jones, M., He, F., Jacobson, A., Pappin, D. J. (December 2004) Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. Mol Cell Proteomics, 3 (12). pp. 1154-69. ISSN 1535-9476 (Print)1535-9476 (Linking)
Abstract
We describe here a multiplexed protein quantitation strategy that provides relative and absolute measurements of proteins in complex mixtures. At the core of this methodology is a multiplexed set of isobaric reagents that yield amine-derivatized peptides. The derivatized peptides are indistinguishable in MS, but exhibit intense low-mass MS/MS signature ions that support quantitation. In this study, we have examined the global protein expression of a wild-type yeast strain and the isogenic upf1Delta and xrn1Delta mutant strains that are defective in the nonsense-mediated mRNA decay and the general 5' to 3' decay pathways, respectively. We also demonstrate the use of 4-fold multiplexing to enable relative protein measurements simultaneously with determination of absolute levels of a target protein using synthetic isobaric peptide standards. We find that inactivation of Upf1p and Xrn1p causes common as well as unique effects on protein expression.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Cations Chromatography, Ion Exchange Chromatography, Liquid Down-Regulation Exoribonucleases/metabolism Fungal Proteins/chemistry Indicators and Reagents/pharmacology Ions Mass Spectrometry Models, Chemical Peptides/chemistry Phenotype Proteomics/ methods RNA Helicases/metabolism RNA, Messenger/metabolism Saccharomyces cerevisiae/ metabolism Saccharomyces cerevisiae Proteins/ chemistry/metabolism Succinimides/chemistry |
| Subjects: | Investigative techniques and equipment bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification organism description > bacteria Investigative techniques and equipment > spectroscopy > mass spectrometry bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein characterization Investigative techniques and equipment > spectroscopy |
| CSHL Authors: | |
| Communities: | CSHL labs > Pappin lab |
| Depositing User: | Matt Covey |
| Date: | December 2004 |
| Date Deposited: | 04 Mar 2013 20:54 |
| Last Modified: | 04 Mar 2013 20:54 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/27713 |
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