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

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 rel...

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Published inJournal of the American Chemical Society Vol. 132; no. 51; pp. 18351 - 18360
Main Authors Szychowski, Janek, Mahdavi, Alborz, Hodas, Jennifer J. L, Bagert, John D, Ngo, John T, Landgraf, Peter, Dieterich, Daniela C, Schuman, Erin M, Tirrell, David A
Format Journal Article
LanguageEnglish
Published WASHINGTON American Chemical Society 29.12.2010
Amer Chemical Soc
Subjects
Alkynes - chemistry
Azides - chemistry
Biotin - chemistry
Chemistry
Chemistry, Multidisciplinary
Cyclization
Dapsone
Green Fluorescent Proteins - chemistry
Molecular Probes - chemical synthesis
Molecular Probes - chemistry
Physical Sciences
Protein Structure, Secondary
Proteins - chemistry
Science & Technology
REPORTER
ANALOGS
RECOMBINANT PROTEINS
IN-VIVO
SOLID-PHASE
CHEMISTRY
LIGATION
TERMINAL ALKYNES
IDENTIFICATION
PROTEOMES
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Summary: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 Na2S2O4, 2% HOCH2CH2SH, 10% HCO2H, 95% CF3CO2H, 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% HCO2H 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.
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ISSN:0002-7863
1520-5126
1520-5126
DOI:10.1021/ja1083909