Journal article
A novel approach to tag and identify geranylgeranylated proteins
Electrophoresis, Vol.30(20), pp.3598-3606
10/2009
DOI: 10.1002/elps.200900259
PMCID: PMC2855049
PMID: 19784953
Abstract
A recently developed proteomic strategy, the “GG-azide”-labeling approach, is described for the detection and proteomic analysis of geranylgeranylated proteins. This approach involves metabolic incorporation of a synthetic azido-geranylgeranyl analog and chemoselective derivatization of azido-geranylgeranyl-modified proteins by the “click” chemistry, using a tetramethylrhodamine-alkyne. The resulting conjugated proteins can be separated by 1-D or 2-D and pH fractionation, and detected by fluorescence imaging. This method is compatible with downstream LC-MS/MS analysis. Proteomic analysis of conjugated proteins by this approach identified several known geranylgeranylated proteins as well as Rap2c, a novel member of the Ras family. Furthermore, prenylation of progerin in mouse embryonic fibroblast cells was examined using this approach, demonstrating that this strategy can be used to study prenylation of specific proteins. The “GG-azide”-labeling approach provides a new tool for the detection and proteomic analysis of geranylgeranylated proteins, and it can readily be extended to other post-translational modifications.
Details
- Title: Subtitle
- A novel approach to tag and identify geranylgeranylated proteins
- Creators
- Lai N Chan - Departments of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USACourtenay Hart - Molecular Probes-Invitrogen, Eugene, OR, USALea Guo - Molecular Biology Institute, University of California, Los Angeles, CA, USATamara Nyberg - Molecular Probes-Invitrogen, Eugene, OR, USABrandon S.J Davies - Departments of Medicine and Human Genetics, University of California, Los Angeles, CA, USALoren G Fong - Departments of Medicine and Human Genetics, University of California, Los Angeles, CA, USAStephen G Young - Departments of Medicine and Human Genetics, University of California, Los Angeles, CA, USABrian J Agnew - Molecular Probes-Invitrogen, Eugene, OR, USAFuyuhiko Tamanoi - Departments of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA
- Resource Type
- Journal article
- Publication Details
- Electrophoresis, Vol.30(20), pp.3598-3606
- DOI
- 10.1002/elps.200900259
- PMID
- 19784953
- PMCID
- PMC2855049
- NLM abbreviation
- Electrophoresis
- ISSN
- 0173-0835
- eISSN
- 1522-2683
- Language
- English
- Date published
- 10/2009
- Academic Unit
- Fraternal Order of Eagles Diabetes Research Center; Biochemistry and Molecular Biology
- Record Identifier
- 9984024510102771
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