Native conjugation between proteins and [60]fullerene derivatives using SpyTag as a reactive handle

Herein, we report the facile conjugation between proteins and water-soluble [60]fullerene derivatives (DC60) under native conditions using SpyTag as a reactive handle. Water-soluble [60]fullerene derivatives were first prepared via sequential Bingel-Hirsch reaction and clicked with SpyTag to give DC60-SpyTag for native conjugation with proteins by the highly efficient SpyTag-SpyCatcher chemistry. The bioconjugation was confirmed by MALDI-TOF MS spectra and SDS-PAGE analysis. The TEM and UV-vis spectroscopic study further revealed that the DC60 could alter the optical performance and induce aggregation of the target proteins. It thus provides a general and robust method for modifying proteins with C-60 derivatives and could potentially be adapted for native conjugation between proteins and other nonbiological motifs as well. (C) 2020 Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences. Published by Elsevier B.V. All rights reserved.

Automated summary

The study demonstrates facile conjugation between proteins and water-soluble [60]fullerene derivatives (DC60) under native conditions using SpyTag. The bioconjugation was confirmed by MALDI-TOF MS spectra and SDS-PAGE analysis, while TEM and UV-vis spectroscopic study revealed the alteration in optical performance induced by DC60 and aggregation of target proteins. This method provides a general and robust approach for modifying proteins with C-60 derivatives.