N-terminal dual protein functionalization by strain-promoted alkyne–nitrone cycloaddition?

Organic & Biomolecular Chemistry Pub Date: 2013-02-20 DOI: 10.1039/C3OB00043E

Abstract

Strain-promoted alkyne–nitrone cycloadditon (SPANC) was optimized as a versatile strategy for dual functionalization of peptides and proteins. The usefulness of the dual labeling protocol is first exemplified by the simultaneous introduction of a chloroquine and a stearyl moiety, two endosomal escape-improving functional groups, into the cell-penetrating peptide hLF (human lactoferrin). Additionally, we demonstrate that dual labeling of proteins is feasible by combining metal-free and copper-catalyzed click chemistry. First, SPANC is applied to enhanced green fluorescent protein to introduce both biotin and a terminal alkyne. The terminal acetylene then serves as a convenient anchor point for the CuAAC reaction with azido-containing fluorescein, thereby demonstrating the potential of combined SPANC and CuAAC for the straightforward, dual functionalization of proteins.

Graphical abstract: N-terminal dual protein functionalization by strain-promoted alkyne–nitrone cycloaddition
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