Living matter is characterized as operating far out of equilibrium, and most of the cellular non-equilibrium processes are organized in extremely complex networks. A visible demonstration of some networks is morphogenesis, which is the driving force for the formation of patterns in tissue. In this project I aim to systematically study the underlying physical and chemical components of morphogenesis in an artificial system. The main characteristics are coupling of chemical reaction and diffusion times, spatial inhomogeneities and positive or negative feedback loops. To achieve this goal, I will assemble an array of prototype ‘cells’ consisting of monodisperse microscopic gel beads produced using microfluidic devices. Within these gel beads, enzymatic reactions will take place to mimic intracellular signaling processes, while the product of the enzymatic reactions can diffuse from one bead to another, thereby resembling cell-cell communication. The enzymatic reactions chosen for my project are self-catalyzed proteolysis and kinase self-phosphorylation, both of which will generate positive feedback. A negative feedback loop will be introduced by using the peptidase cascade by hydrolysis of a pre-inhibitor, releasing a free inhibitor after cleavage.
An analysis of the reaction diffusion profile is a novel method for measuring enzyme kinetics. As a spin-off from this project, I am interested to use a reaction-diffusion between two drops for the study of interaction of enzyme with inhibitors, in order to miniaturize and accelerate an inhibitor screening.
Field of science
- /natural sciences/biological sciences/biochemistry/biomolecules/proteins/enzymes
Call for proposal
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