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			PubMed Journals: Proc Natl Acad Sci U S A

  Source:		PMID: 15308774


    		Proc Natl Acad Sci U S A. 2004 Aug
     		24;101(34):12479-84. Epub 2004 Aug 12.

			A tagging-via-substrate technology for detection
			and proteomics of farnesylated proteins.

			Kho Y(1), Kim SC, Jiang C, Barma D, Kwon
			SW, Cheng J, Jaunbergs J, Weinbaum C, Tamanoi
			F, Falck J, Zhao Y.

			Author Information
			(1) Department of Biochemistry,
			University of Texas Southwestern Medical Center,
			Dallas, TX 75390-9038.

			A recently developed proteomics strategy,
			designated tagging-via-substrate (TAS) approach,
			is described for the detection and proteomic
			analysis of farnesylated proteins. TAS technology
			involves metabolic incorporation of a synthetic
			azido-farnesyl analog and chemoselective
			derivatization of azido-farnesyl-modified
			proteins by an elegant version of Staudinger
			reaction, pioneered by the Bertozzi group,
			using a biotinylated phosphine capture reagent.
			The resulting protein conjugates can be
			specifically detected and/or affinity-purified
			by streptavidin-linked horseradish peroxidase
			or agarose beads, respectively. Thus, the
			technology enables global profiling of farnesylated
			proteins by enriching farnesylated proteins
			and reducing the complexity of farnesylation
			subproteome. Azido-farnesylated proteins
			maintain the properties of protein farnesylation,
			including promoting membrane association,
			Ras-dependent mitogen-activated protein
			kinase kinase activation, and inhibition
			of lovastatin-induced apoptosis. A proteomic
			analysis of farnesylated proteins by TAS
			technology revealed 18 farnesylated proteins,
			including those with potentially novel farnesylation
			motifs, suggesting that future use of this
			method is likely to yield novel insight
			into protein farnesylation. TAS technology
			can be extended to other posttranslational
			modifications, such as geranylgeranylation
			and myristoylation, thus providing powerful
			tools for detection, quantification, and
			proteomic analysis of posttranslationally
			modified proteins.

			DOI: 10.1073/pnas.0403413101 PMCID: PMC515085
			PMID: 15308774 [Indexed for MEDLINE]

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