@ARTICLE{Aper2018-qv,
  title    = "{Protease-Activatable} Scaffold Proteins as Versatile Molecular
              Hubs in Synthetic Signaling Networks",
  author   = "Aper, Stijn J A and den Hamer, Anniek and Wouters, Simone F A and
              Lemmens, Lenne J M and Ottmann, Christian and Brunsveld, Luc and
              Merkx, Maarten",
  abstract = "Protease signaling and scaffold-induced control of
              protein-protein interactions represent two important mechanisms
              for intracellular signaling. Here we report a generic and modular
              approach to control the activity of scaffolding proteins by
              protease activity, creating versatile molecular platforms to
              construct synthetic signaling networks. Using 14-3-3 proteins as
              a structurally well-characterized and important class of scaffold
              proteins, three different architectures were explored to achieve
              optimal protease-mediated control of scaffold activity, fusing
              either one or two monovalent inhibitory ExoS peptides or a single
              bivalent ExoS peptide to T14-3-3 using protease-cleavable
              linkers. Analysis of scaffolding activity before and after
              protease-induced cleavage revealed optimal control of 14-3-3
              activity for the system that contained monovalent ExoS peptides
              fused to both the N-and C-terminus, each blocking a single
              T14-3-3 binding site. The protease-activatable 14-3-3 scaffolds
              were successfully applied to construct a three-step signaling
              cascade in which dimerization and activation of FGG-caspase-9 on
              an orthogonal supramolecular platform resulted in activation of a
              14-3-3 scaffold, which in turn allowed 14-3-3-templated
              complementation of a split-luciferase. In addition, by combining
              14-3-3-templated activation of caspase-9 with a
              caspase-9-activatable 14-3-3 scaffold, the first example of a
              synthetic self-activating protease signaling network was created.
              Protease-activatable 14-3-3 proteins thus represent a modular
              platform whose properties can be rationally engineered to fit
              different applications, both to create artificial in vitro
              synthetic molecular networks and as a novel signaling hub to
              re-engineer intracellular signaling pathways.",
  journal  = "ACS Synth Biol",
  volume   =  7,
  number   =  9,
  pages    = "2216--2225",
  month    =  aug,
  year     =  2018,
  keywords = "14-3-3 protein; proteases; protein engineering; self-activation;
              signaling cascade; synthetic biology",
  language = "en"
}

{"_id":"ckZgQ5kHYNyHHBAcQ","bibbaseid":"aper-denhamer-wouters-lemmens-ottmann-brunsveld-merkx-proteaseactivatablescaffoldproteinsasversatilemolecularhubsinsyntheticsignalingnetworks-2018","author_short":["Aper, S. J A","den Hamer, A.","Wouters, S. F A","Lemmens, L. J M","Ottmann, C.","Brunsveld, L.","Merkx, M."],"bibdata":{"bibtype":"article","type":"article","title":"Protease-Activatable Scaffold Proteins as Versatile Molecular Hubs in Synthetic Signaling Networks","author":[{"propositions":[],"lastnames":["Aper"],"firstnames":["Stijn","J","A"],"suffixes":[]},{"propositions":["den"],"lastnames":["Hamer"],"firstnames":["Anniek"],"suffixes":[]},{"propositions":[],"lastnames":["Wouters"],"firstnames":["Simone","F","A"],"suffixes":[]},{"propositions":[],"lastnames":["Lemmens"],"firstnames":["Lenne","J","M"],"suffixes":[]},{"propositions":[],"lastnames":["Ottmann"],"firstnames":["Christian"],"suffixes":[]},{"propositions":[],"lastnames":["Brunsveld"],"firstnames":["Luc"],"suffixes":[]},{"propositions":[],"lastnames":["Merkx"],"firstnames":["Maarten"],"suffixes":[]}],"abstract":"Protease signaling and scaffold-induced control of protein-protein interactions represent two important mechanisms for intracellular signaling. Here we report a generic and modular approach to control the activity of scaffolding proteins by protease activity, creating versatile molecular platforms to construct synthetic signaling networks. Using 14-3-3 proteins as a structurally well-characterized and important class of scaffold proteins, three different architectures were explored to achieve optimal protease-mediated control of scaffold activity, fusing either one or two monovalent inhibitory ExoS peptides or a single bivalent ExoS peptide to T14-3-3 using protease-cleavable linkers. Analysis of scaffolding activity before and after protease-induced cleavage revealed optimal control of 14-3-3 activity for the system that contained monovalent ExoS peptides fused to both the N-and C-terminus, each blocking a single T14-3-3 binding site. The protease-activatable 14-3-3 scaffolds were successfully applied to construct a three-step signaling cascade in which dimerization and activation of FGG-caspase-9 on an orthogonal supramolecular platform resulted in activation of a 14-3-3 scaffold, which in turn allowed 14-3-3-templated complementation of a split-luciferase. In addition, by combining 14-3-3-templated activation of caspase-9 with a caspase-9-activatable 14-3-3 scaffold, the first example of a synthetic self-activating protease signaling network was created. Protease-activatable 14-3-3 proteins thus represent a modular platform whose properties can be rationally engineered to fit different applications, both to create artificial in vitro synthetic molecular networks and as a novel signaling hub to re-engineer intracellular signaling pathways.","journal":"ACS Synth Biol","volume":"7","number":"9","pages":"2216–2225","month":"August","year":"2018","keywords":"14-3-3 protein; proteases; protein engineering; self-activation; signaling cascade; synthetic biology","language":"en","bibtex":"@ARTICLE{Aper2018-qv,\n title = \"{Protease-Activatable} Scaffold Proteins as Versatile Molecular\n Hubs in Synthetic Signaling Networks\",\n author = \"Aper, Stijn J A and den Hamer, Anniek and Wouters, Simone F A and\n Lemmens, Lenne J M and Ottmann, Christian and Brunsveld, Luc and\n Merkx, Maarten\",\n abstract = \"Protease signaling and scaffold-induced control of\n protein-protein interactions represent two important mechanisms\n for intracellular signaling. Here we report a generic and modular\n approach to control the activity of scaffolding proteins by\n protease activity, creating versatile molecular platforms to\n construct synthetic signaling networks. Using 14-3-3 proteins as\n a structurally well-characterized and important class of scaffold\n proteins, three different architectures were explored to achieve\n optimal protease-mediated control of scaffold activity, fusing\n either one or two monovalent inhibitory ExoS peptides or a single\n bivalent ExoS peptide to T14-3-3 using protease-cleavable\n linkers. Analysis of scaffolding activity before and after\n protease-induced cleavage revealed optimal control of 14-3-3\n activity for the system that contained monovalent ExoS peptides\n fused to both the N-and C-terminus, each blocking a single\n T14-3-3 binding site. The protease-activatable 14-3-3 scaffolds\n were successfully applied to construct a three-step signaling\n cascade in which dimerization and activation of FGG-caspase-9 on\n an orthogonal supramolecular platform resulted in activation of a\n 14-3-3 scaffold, which in turn allowed 14-3-3-templated\n complementation of a split-luciferase. In addition, by combining\n 14-3-3-templated activation of caspase-9 with a\n caspase-9-activatable 14-3-3 scaffold, the first example of a\n synthetic self-activating protease signaling network was created.\n Protease-activatable 14-3-3 proteins thus represent a modular\n platform whose properties can be rationally engineered to fit\n different applications, both to create artificial in vitro\n synthetic molecular networks and as a novel signaling hub to\n re-engineer intracellular signaling pathways.\",\n journal = \"ACS Synth Biol\",\n volume = 7,\n number = 9,\n pages = \"2216--2225\",\n month = aug,\n year = 2018,\n keywords = \"14-3-3 protein; proteases; protein engineering; self-activation;\n signaling cascade; synthetic biology\",\n language = \"en\"\n}\n\n","author_short":["Aper, S. J A","den Hamer, A.","Wouters, S. F A","Lemmens, L. J M","Ottmann, C.","Brunsveld, L.","Merkx, M."],"key":"Aper2018-qv","id":"Aper2018-qv","bibbaseid":"aper-denhamer-wouters-lemmens-ottmann-brunsveld-merkx-proteaseactivatablescaffoldproteinsasversatilemolecularhubsinsyntheticsignalingnetworks-2018","role":"author","urls":{},"keyword":["14-3-3 protein; proteases; protein engineering; self-activation; signaling cascade; synthetic biology"],"metadata":{"authorlinks":{}}},"bibtype":"article","biburl":"https://bibbase.org/network/files/YGLkdBTPJKsfXw8eh","dataSources":["cxRx9FFoorWFRTNbm","gQRZwEYHHnkKBfPxS","KxmF4kcmdXDANWdBG","6WDRpfi2GcduYyzsq","ba7dRGjRxop47uYRZ"],"keywords":["14-3-3 protein; proteases; protein engineering; self-activation; signaling cascade; synthetic biology"],"search_terms":["protease","activatable","scaffold","proteins","versatile","molecular","hubs","synthetic","signaling","networks","aper","den hamer","wouters","lemmens","ottmann","brunsveld","merkx"],"title":"Protease-Activatable Scaffold Proteins as Versatile Molecular Hubs in Synthetic Signaling Networks","year":2018}