Funding
Our research is made possible by funding from:
Living systems are highly dynamic, continuously reshaping and reorganising as their molecular components associate and dissociate. Rather than relying on deterministic genetic programmes, intermolecular interactions stay flexible, enabling them to respond to environmental changes and maintain robust cell response. Our research aims to elucidate how ensemble properties of the cellular interface and functions of individual molecules are modulated by spatial confinement or the presence of regulatory elements, such as cofactors or inhibitors. We also explore how other cells recognise such controlled mimetic interfaces.
Although inherently reductionist, the bottom-up reconstitution approach is uniquely positioned to dissect the complexity of these interactions and bridge the gap between molecular and cell-level organisation. Our primary focus is on reconstructing glycan–protein assemblies associated with lipid membranes, which we characterise using a range of surface-sensitive techniques. Whenever possible, we perform “biochemistry under the microscope,” directly observing and quantifying dynamic interactions by single-molecule tracking (video on the right).
Peptidoglycan synthesis
While enzymes involved in peptidoglycan synthesis are popular targets for antibiotics, their activity largely depends on the cellular context and is controlled by different molecular regulators. We rebuild molecular networks that control peptidoglycan synthesis during bacterial division. By accounting on relevant boundary conditions, such as confinement to cellular membranes, presence of peptidoglycan or FtsZ cytoskeleton we can quantify dynamics of the interactions with a control and precision that remains difficult to achieve in living cell. Our reconstitution assays can be also adapted as high-throughput screening platform for target-based discovery of novel peptidoglycan synthesis inhibitors.
Bacteria-host communication
Biofilm matrix, where glycans represent one of the main structural components, serves as communication media between bacterial cells and other species. While the chemical means of communication, such as quorum sensing, received decades of attention, how bacteria react to the change in composition and mechanical remodeling of their own matrix or the matrix produced by another organism is far less understood. We aim to rebuild biofilm matrix components on a planar surface to investigate molecular interactions and monitor its recognition by bacterial cells or host immune cells at high spatio-temporal resolution. Another objective is to investigate how environment of the host tissue affects bacterial pathogenesis and biofilm formation.
Dynamics of glycocalyx
Bulky glycocalyx components are osmotically active space fillers that define the thickness and viscoelastic properties of the cell interface. However, the relation between their ensemble architecture, dynamic re-organization and cell functions remains largely unexplored for most cells. Here, we focus on the development of novel approaches to probe physical properties of the glycocalyx in synthetic assembly and correlat with the glycocalyx assembly present on a living cell.
