Microfluidic Analysis of Multicomponent Phase Separation in Ubiquitin-Dependent Proteostasis
We develop microfluidic workflows to map enzymatic activity and compositional control of tumour suppressor SPOP with its substrates in liquid nuclear bodies. Various membraneless bodies that form through liquid phase separation have been described in recent years. While many components of liquid phase separated bodies were identified and successfully reconstituted in vitro, general principles that define body composition and function are difficult to characterise experimentally. This is in particular the case for dynamic aspects, such as early and later stages or hysteresis in liquid-liquid phase separation, molecular ordering and reorganisation dynamics, as well as quantification of enzymatic activities within phase separated bodies. Furthermore, several phase-separated bodies may co-exist and enzymes and their substrates may partition into multiple bodies, posing questions as to how such redistribution between bodies affects function and how it is regulated.
The ‘Intelligent Biointegrative Systems’ lab develops, tests and validates new biobased and biocompatible materials and processes for the application of generative manufacturing processes in life sciences in an interdisciplinary approach. To this end, the building blocks of life (proteins, lipid membranes and DNA) are structured into three-dimensional scaffolds and complex reaction spaces. All relevant length scales from the molecular scale to the cell and tissue level are integrated and combined with self-assembly and lithographic methods. Our long-term goal is to achieve the most detailed (re-)construction of cellular structures and functional elements with the highest possible precision and controllability to advance our understandig of biomolecular and cell biological processes. This enables revolutionary new biomolecular structures and functions for applications in pharmaceutical, biotech and biomedical research, as well as in the production of tissue substitutes. In addition to new design methods for biomolecular process engineering, we are also developing new innovative methods for time-resolved sequence-structure-function analysis of biomolecular dynamics.
In particular working on the project: Tanja Mittag, St. Jude Children’s Research Hospital