title: Ultrasensitivity by molecular titration in spatially propagating enzymatic reactions.
abstract: Delineating design principles of biological systems by reconstitution of purified components offers a platform to gauge the influence of critical physicochemical parameters on minimal biological systems of reduced complexity. Here we unravel the effect of strong reversible inhibitors on the spatiotemporal propagation of enzymatic reactions in a confined environment in vitro. We use micropatterned, enzyme-laden agarose gels which are stamped on polyacrylamide films containing immobilized substrates and reversible inhibitors. Quantitative fluorescence imaging combined with detailed numerical simulations of the reaction-diffusion process reveal that a shallow gradient of enzyme is converted into a steep product gradient by addition of strong inhibitors, consistent with a mathematical model of molecular titration. The results confirm that ultrasensitive and threshold effects at the molecular level can convert a graded input signal to a steep spatial response at macroscopic length scales.
authors: Radboud University Nijmegen, Institute for Molecules and Materials, Nijmegen, The Netherlands.
authors: Sergey N
authors: SN
authors: Semenov
authors: Albert J
authors: AJ
authors: Markvoort
authors: Wouter B L
authors: WB
authors: Gevers
authors: Aigars
authors: A
authors: Piruska
authors: Tom F A
authors: TF
authors: de Greef
authors: Wilhelm T S
authors: WT
authors: Huck
mesh_terms: D004791:Enzyme Inhibitors / Q000494:pharmacology; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D008970:Molecular Weight; D015203:Reproducibility of Results; D062211:Spatio-Temporal Analysis
chemical_list: D004791:Enzyme Inhibitors; D004798:Enzymes
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