Fig. 1: The NeissLock concept and optimizing autoproteolysis.
From: NeissLock provides an inducible protein anhydride for covalent targeting of endogenous proteins
a The FrpC self-processing module (SPM) catalyzes autoproteolytic cleavage at an Asp-Pro peptide bond, induced by calcium, generating an anhydride. b NeissLock vision. SPM (orange) is recombinantly fused to a binding protein (blue) which docks with the target protein (green). Adding calcium promotes generation of the anhydride (yellow star), so the binding protein can form a covalent bond to the target protein. c Schematic of reaction of amine on target protein with aspartic anhydride on binding protein. d Time-course of SPM cleavage with Ala preceding Asp-Pro, analyzed by SDS-PAGE with Coomassie staining (pH 7.4, 37 °C). Molecular weight markers represent kDa. e Histogram of SPM cleavage rate with each residue before Asp-Pro, moving from the least cleaved residue at 15 min on the left to the most cleaved residue on the right (mean of triplicate ± 1 s.d.; some error bars are too small to be visible; pH 7.4 and 37 °C. Source data are provided as a Source Data file.
