A Highly Efficient Method for Site-Specific Modification of Unprotected Peptides after Chemical Synthesis
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- 作者：Steven J. Bark ; Sandra Schmid ; and Klaus M. Hahn
- 刊名：Journal of the American Chemical Society
- 出版年：2000
- 出版时间：April 19, 2000
- 年：2000
- 卷：122
- 期：15
- 页码：3567 - 3573
- 全文大小：133K
- 年卷期：v.122,no.15(April 19, 2000)
- ISSN：1520-5126
文摘
We have developed a highly efficient method for the site-specific attachment of biophysical probesto unprotected peptides after chemical synthesis. This methodology takes advantage of the selective reactivityof an N-methylaminooxy amino acid that is appropriately protected for direct incorporation during solid-phasepeptide synthesis. The functional N-methylaminooxy group is unmasked using normal peptide cleavageconditions and is capable of selective reaction with activated N-hydroxysuccinimide esters in the presence ofcysteine, lysine and the amino terminus, as demonstrated in model peptides and test proteins. Selective labelingcan be accomplished after synthesis using commercially available or chemically sensitive probes. This technologyis compatible with the synthesis of C
-thioester-containing peptides and amide-forming ligations, requiredsteps for the synthesis of proteins by either total chemical synthesis or expressed protein ligation. TheN-methylaminooxy amino acid can be introduced into different sites by parallel peptide synthesis to generatea polypeptide analogue family with each member possessing a single specifically labeled site. This enablesthe synthesis of optimized biosensors through combinatorial screening of different attachment sites for maximalresponse and minimal perturbation of biological activity.
-thioester-containing peptides and amide-forming ligations, requiredsteps for the synthesis of proteins by either total chemical synthesis or expressed protein ligation. TheN-methylaminooxy amino acid can be introduced into different sites by parallel peptide synthesis to generatea polypeptide analogue family with each member possessing a single specifically labeled site. This enablesthe synthesis of optimized biosensors through combinatorial screening of different attachment sites for maximalresponse and minimal perturbation of biological activity.