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. Author manuscript; available in PMC: 2021 Jun 7.
Published in final edited form as: Nat Chem Biol. 2020 Dec 7;17(3):272–279. doi: 10.1038/s41589-020-00696-0

Extended Data Fig. 1. UBE2N~UB/UBE2V1/RNF4 RING E3 complex reacts preferentially with free amino acids harboring amine acceptors and various side-chain.

Extended Data Fig. 1

(a) Fluorescence scan of SDS-PAGE gels demonstrating the discharge of labeled UB (UB*) to L-lysine compared with the absence of amino acid acceptor using wild-type UBE2N. Electrophoresis was performed under both reducing and non-reducing conditions to differentiate thioester bonded complexes from isopeptide bonded E2-donor UB ones. (b) Time-courses of fluorescent UB discharge from UBE2N K92R~UB/UBE2V1/RNF4 RING E3 to the indicated amino acids, normalized to starting signal of fluorescent UB thioester-bonded to UBE2N. For all, N=2 independent experiments. For samples derived from the same experiment, gels were processed in parallel.