Protein labeling and crosslinking by covalent aptamers

In this chapter, a new approach to the selective modification of native proteins is discussed, using electrophilic covalent aptamers. These biochemical tools are generated through the site-specific incorporation of a label-transferring or crosslinking electrophile into a DNA aptamer. Covalent aptame...

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Veröffentlicht in:Methods in enzymology 2023-01, Vol.682, p.413-428
Hauptverfasser: Cacace, Mary, Tivon, Yaniv, Deiters, Alexander
Format: Artikel
Sprache:eng
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Zusammenfassung:In this chapter, a new approach to the selective modification of native proteins is discussed, using electrophilic covalent aptamers. These biochemical tools are generated through the site-specific incorporation of a label-transferring or crosslinking electrophile into a DNA aptamer. Covalent aptamers provide the ability to transfer a variety of functional handles to a protein of interest or to irreversibly crosslink to the target. Methods for the aptamer-mediated labeling and crosslinking of thrombin are described. Thrombin labeling is fast and selective, in both simple buffer and in human plasma and outcompetes nuclease-mediated degradation. This approach provides facile, sensitive detection of labeled protein by western blot, SDS-PAGE, and mass spectrometry.
ISSN:0076-6879
1557-7988
DOI:10.1016/bs.mie.2022.08.053