Biologically Inspired Molecular Adhesives towards Multifunctional Biomaterials and Microreactors

Projektinformationen

VELCROProtein

ID Finanzhilfevereinbarung: 101002094

DOI

10.3030/101002094

EK-Unterschriftsdatum 20 Januar 2021

Startdatum 1 September 2021

Enddatum 31 August 2026

Finanziert unter

• EXCELLENT SCIENCE - European Research Council (ERC)

Gesamtkosten

€ 1 998 496,00

EU-Beitrag

€ 1 998 496,00

Koordiniert durch

EIDGENOESSISCHE TECHNISCHE HOCHSCHULE ZUERICH
Switzerland

Periodic Reporting for period 2 - VELCROProtein (Biologically Inspired Molecular Adhesives towards Multifunctional Biomaterials and Microreactors)

Berichtszeitraum: 2023-03-01 bis 2024-08-31

Zusammenfassung vom Kontext und den Gesamtzielen des Projekts

The human body is composed of biomolecules such as proteins and nucleic acids, which interact spontaneously to form various supramolecular structures, including compartments and fibrils. The properties of these structures, such as size and shape, depend on both the information encoded in the molecular sequence and the local environment. This innate ability of biological molecules to self-organize into functional structures lies at the core of life. Indeed, these structures coordinate the myriad biochemical activities necessary for cellular survival, growth, and reproduction. Much like factories, cells must efficiently coordinate numerous biochemical reactions in both space and time. Compartmentalization serves as a key strategy to achieve this coordination. Traditionally, biological studies have focused on membrane-bound compartments, where physical barriers separate them from the surrounding environment. However, recent research has revealed the existence of membraneless compartments, known as biomolecular condensates, which form through the phase separation of proteins and nucleic acids. This discovery has opened many questions regarding the molecular mechanisms governing the formation and properties of membraneless compartments, as well as the interplay between these compartments and biochemical reactions. This project has two primary objectives: a) to generate simplified models of biological membraneless compartments in vitro, allowing for the investigation of the physicochemical principles underlying their properties and their link with the modulation of biochemical reactions; b) to leverage this understanding to design novel molecules with programmable phase separation behavior which do no exist in nature. Special focus is given on studying enzymatic reactions and aggregation events within open compartments.

Arbeit, die ab Beginn des Projekts bis zum Ende des durch den Bericht erfassten Berichtszeitraums geleistet wurde, und die wichtigsten bis dahin erzielten Ergebnisse

In this first part of the project, we have succesfully accomplished all planned goals. We have produced various biological proteins associated with the formation of biological compartments and studied their behavior through a range of biophysical techniques. This approach has enabled us to unravel the intricate relationship between the sequence of the molecules and the properties of the compartments. Since cells do not operate at thermodynamic equilibrium but rather at steady states, our focus has been on analysising the behavior of the compartments over time. Through this investigation, we have identified key properties of the compartments that regulate biochemical activities, such as enzymatic reactions and aggregation events. Notably, we have identified the crucial role of the interface and of the composition of the compartments in regulating these acitivities. Moreover, we have developed new tools based on microfluidic technology to accurately investigate phase separation, achieving precise control over all relevant biochemical and biophysical parameters.
Based on the lessons learnt from biologival molecules, we have engineered synthetic proteins capable of forming compartments with tailored phase separation behaviors. Specifically, we have generated compartments capable of modulating enzymatic reactions, uncovering design principles that enhance reaction performance.

Fortschritte, die über den aktuellen Stand der Technik hinausgehen und voraussichtliche potenzielle Auswirkungen (einschließlich der bis dato erzielten sozioökonomischen Auswirkungen und weiter gefassten gesellschaftlichen Auswirkungen des Projekts)

In some cases, the complexity of biological systems cannot be fully understood by studying living organisms directly. Reproducing simplified models of biological compartments in test tubes represents a powerful strategy to complement in vivo studies and elucidate the physicochemical rules governing the phase separation behavior of proteins and nucleic acids. Our results show the complex nature of the phase separation of these molecules, which is regulated by a delicate balance of multiple factors. Understanding these governing principles holds significant implications in several directions.
First of all, it aids in identifying mechanisms that control the formation of biological compartments within cells. This is particuarly crucial in scenarios where intervention is necessary to rectify improper behavior. A notable example is the mis-regulation of certain compartments associated with age-related diseases such neurodegenerative disorders. By understanding the mechanisms regulating functional and dysfunctional behavior, we may one day develop strategies to correct aberrant events, such as the undesired formation of protein aggregates within compartments, which has been linked to pathology. Morever, in addition to contributions to biological studies, the results of this project have implications in the design of novel advanced materials based on programmable phase separation. By learning lessons from cellular processes, we can engineer proteins and nucleic acids to form tailored compartments capable of regulating biochemical reactions or releasing molecules on demand. These materials hold promise for diverse applications across various fields, including synthetic biology, the manufacturing of biological drugs, diagnostics, and drug delivery. Some of these applications have already been explored in the first half of this project in our laboratory.

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