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Science 16 January 2004:
Vol. 303. no. 5656, pp. 371 - 373
DOI: 10.1126/science.1089509
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Reports
A New Strategy for the Synthesis of Glycoproteins
Zhiwen Zhang,1
Jeff Gildersleeve,1
Yu-Ying Yang,1
Ran Xu,1
Joseph A. Loo,2
Sean Uryu,1
Chi-Huey Wong,1
Peter G. Schultz1*
Posttranslational modifications of proteins regulate many biological processes, including metabolism, signal transduction, and gene expression. The synthetic challenges associated with generating homogeneous populations of selectively modified proteins, however, have hindered detailed studies of the effects of these modifications on protein structure and function. Here, we report an approach to the cotranslational synthesis of selectively glycosylated proteins in which the modified amino acid is genetically encoded. We show that myoglobin containing ß–N-acetylglucosamine (GlcNAc)–serine at a defined position can be expressed in Escherichia coli in good yield and with high fidelity. The ß-GlcNAc moiety can be recognized by a saccharide-binding protein, or subsequently modified with a galactosyltransferase to build more complex carbohydrates. This approach should be generally applicable to other posttranslational modifications such as protein phosphorylation, acetylation, and methylation.
1 Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
2 Department of Biochemistry and Biological Chemistry, University of California at Los Angeles, 405 Hilgard Avenue, 402 Paul D. Boyer Hall, Los Angeles, CA 90095, USA.
* To whom correspondence should be addressed. E-mail: schultz{at}scripps.edu
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