Extended Data Fig. 2: SDS-PAGE analysis of in-solution treatment of autonomously compiled mGreenLantern pendants.
From: Boolean logic-gated protein presentation through autonomously compiled molecular topology
a, mGreenLantern-A∧(S∨T) is differentially cleaved following the 8 possible input combinations of A, S, and T. Input conditions A, S, T, and ST are expected to linearize the product but not induce payload release, leading to a product with less electrophoretic mobility than the initial cyclic construct. Payload release is expected following input conditions AS, AT, and AST, accompanied with a product band that migrates further than the starting species. b, Changes in topology and/or molecular weight in response to a specific set of inputs leads to changes in protein electrophoretic mobility, which can be analyzed through gel densitometry. c, The response profiles of the YES-gated single-input tethers. d-e, The response profiles of the two-input (d) OR-gated and (e) AND-gated tethers. f-h, The response profiles of the three-input (f) OR/(AND)-, (g) AND/(OR)-, and (h) OR/OR-gated tethers. Plot titles correspond to the protein tether identity; the y-axis represents extent of cleavage as measured through migration on an SDS-PAGE with gel densitometry analysis; the x-axis indicates treatment conditions, wherein N indicates no treatment, A indicates eSrtA(2A9), S indicates eSrtA(4S9), and T indicates TEV. Green bars indicate conditions expected to yield tether cleavage, whereas red bars indicate conditions expected to keep the tether non-cleaved. Error bars correspond to ±1 standard deviation about the mean with propagated uncertainties for n = 3 experimental replicates.
