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Cleavable Biotin Probes for Labeling of Biomolecules via Azide−Alkyne Cycloaddition

Szychowski, Janek and Mahdavi, Alborz and Hodas, Jennifer J. L. and Bagert, John D. and Ngo, John T. and Landgraf, Peter and Dieterich, Daniela C. and Schuman, Erin M. and Tirrell, David A. (2010) Cleavable Biotin Probes for Labeling of Biomolecules via Azide−Alkyne Cycloaddition. Journal of the American Chemical Society, 132 (51). pp. 18351-18360. ISSN 0002-7863. PMCID PMC3016050. http://resolver.caltech.edu/CaltechAUTHORS:20110303-113837195

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Abstract

The azide−alkyne cycloaddition provides a powerful tool for bio-orthogonal labeling of proteins, nucleic acids, glycans, and lipids. In some labeling experiments, e.g., in proteomic studies involving affinity purification and mass spectrometry, it is convenient to use cleavable probes that allow release of labeled biomolecules under mild conditions. Five cleavable biotin probes are described for use in labeling of proteins and other biomolecules via azide−alkyne cycloaddition. Subsequent to conjugation with metabolically labeled protein, these probes are subject to cleavage with either 50 mM Na_2S_2O_4, 2% HOCH_2CH_2SH, 10% HCO_2H, 95% CF_3CO_2H, or irradiation at 365 nm. Most strikingly, a probe constructed around a dialkoxydiphenylsilane (DADPS) linker was found to be cleaved efficiently when treated with 10% HCO_2H for 0.5 h. A model green fluorescent protein was used to demonstrate that the DADPS probe undergoes highly selective conjugation and leaves a small (143 Da) mass tag on the labeled protein after cleavage. These features make the DADPS probe especially attractive for use in biomolecular labeling and proteomic studies.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ja1083909 DOIArticle
http://pubs.acs.org/doi/abs/10.1021/ja1083909PublisherArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3016050/PubMed CentralArticle
ORCID:
AuthorORCID
Mahdavi, Alborz0000-0002-8790-8112
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2010 American Chemical Society. Received September 16, 2010, Publication Date (Web): December 8, 2010. This work was supported by NIH GM62523 (DAT), NIH MH065537 (EMS), the ARO Institute for Collaborative Biotechnologies, and Fonds Québécois de la Recherche sur la Nature et les Technologies (as a postdoctoral fellowship to J.S.). A.M. was supported in part by a National Science and Engineering Council of Canada (NSERC) postgraduate scholarship. We thank Sonja Hess, Jacob Bitterman, Eva Wurster, and Christiane Kowatsch for helpful discussions, as well as the Proteome Exploration Laboratory (PEL), MS, and NMR facilities of the California Institute of Technology. Sequence and mass spectrum of protein 18; ^1H NMR spectra for compounds 4, 5, 6, 9, 11, 12, 14a, and 14b; ^(13)C NMR spectra for compounds 4, 5, 6, 9, 12, 14a, and 14b; MS/MS spectra of tryptic fragments.
Funders:
Funding AgencyGrant Number
NIHGM62523
NIHMH065537
Army Research Office (ARO)UNSPECIFIED
Fonds Québécois de la Recherche sur la Nature et les Technologies (FQRNT)UNSPECIFIED
Natural Sciences and Engineering Research Council of Canada (NSERC)UNSPECIFIED
PubMed Central ID:PMC3016050
Record Number:CaltechAUTHORS:20110303-113837195
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20110303-113837195
Official Citation:Cleavable Biotin Probes for Labeling of Biomolecules via Azide−Alkyne Cycloaddition Janek Szychowski, Alborz Mahdavi, Jennifer J. L. Hodas, John D. Bagert, John T. Ngo, Peter Landgraf, Daniela C. Dieterich, Erin M. Schuman, David A. Tirrell Journal of the American Chemical Society 2010 132 (51), 18351-18360
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:22639
Collection:CaltechAUTHORS
Deposited By: Benjamin Perez
Deposited On:03 Mar 2011 23:09
Last Modified:05 Jan 2016 21:12

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