Skip to main content
. Author manuscript; available in PMC: 2015 Jul 21.
Published in final edited form as: Chembiochem. 2014 Jun 24;15(11):1563–1568. doi: 10.1002/cbic.201402059

Scheme 1.

Scheme 1

Synthesis of dually functionalized Ub polymers by exploiting the transamidase activity of Yuh1. (A) Scheme depicting our previous work using Yuh1 to replace D77 of Ub with allylamine and subsequent formation of Ub polymers via thiol-ene coupling (TEC). The linkage afforded by TEC is a functional mimic of the native isopeptide bond. For the purpose of differentiating between native isopeptide bonds and TEC-derived non-native linkages, “Kx-linked” represents the former and “x-linked” represents the latter where×is any of the seven native lysine positions. (B) Scheme showing the studies described here in which Yuh1 catalyzes the C-terminal installation of amine-biotin I. The polymers are subsequently tagged with a fluorophore (blue star) and immobilized on a polyethylene glycol (PEG)-modified glass surface for detection by SMF.