Periodic Reporting for period 1 - Pressure-Jump NMR (Investigation of aggregation-prone and phase-separating systems using Pressure-Jump NMR)
Periodo di rendicontazione: 2023-09-01 al 2025-08-31
Proteins are the molecular machines that sustain life. They fold into precise shapes to perform their biological functions, but when this process goes wrong, proteins can clump together and form aggregates. Such aggregation underlies many serious diseases, including Alzheimer’s, Parkinson’s, and type II diabetes. Despite decades of research, we still struggle to observe and understand how proteins aggregate and separate into different liquid phases inside cells, because these processes are extremely fast and difficult to capture experimentally.
The Pressure-Jump NMR (PJ-NMR) project set out to establish a completely new way of studying these dynamic protein behaviors. Using controlled bursts of pressure, PJ-NMR allows us to trigger and observe molecular changes as they happen, at the level of individual atoms. This new experimental approach promises to reveal details of how proteins interact, fold, and form aggregates or droplets — processes central to many biological functions and disease mechanisms.
The project’s main objectives were to:
-Design, build, and validate a new PJ-NMR instrument in Europe.
-Develop new measurement methods to capture the rapid molecular events following a pressure change.
-Apply these methods to study proteins involved in aggregation and liquid–liquid phase separation.
-Lay the foundation for a sustainable European research platform capable of using PJ-NMR to tackle urgent biomedical challenges.
By advancing both instrumentation and methodology, the project addresses a major scientific need: to connect the static structural information available from conventional techniques with the dynamic, real-time processes that drive disease and biological function. In the long term, this technology will contribute to Europe’s capacity to study, diagnose, and eventually control protein misfolding diseases — an area of increasing health and economic significance given the ageing population.
The Pressure-Jump NMR (PJ-NMR) project set out to establish a completely new way of studying these dynamic protein behaviors. Using controlled bursts of pressure, PJ-NMR allows us to trigger and observe molecular changes as they happen, at the level of individual atoms. This new experimental approach promises to reveal details of how proteins interact, fold, and form aggregates or droplets — processes central to many biological functions and disease mechanisms.
The project’s main objectives were to:
-Design, build, and validate a new PJ-NMR instrument in Europe.
-Develop new measurement methods to capture the rapid molecular events following a pressure change.
-Apply these methods to study proteins involved in aggregation and liquid–liquid phase separation.
-Lay the foundation for a sustainable European research platform capable of using PJ-NMR to tackle urgent biomedical challenges.
By advancing both instrumentation and methodology, the project addresses a major scientific need: to connect the static structural information available from conventional techniques with the dynamic, real-time processes that drive disease and biological function. In the long term, this technology will contribute to Europe’s capacity to study, diagnose, and eventually control protein misfolding diseases — an area of increasing health and economic significance given the ageing population.
During the fellowship, a fully functional PJ-NMR instrument was designed, constructed, and successfully implemented at Lund University (Sweden). This represents the first European installation of such a device, making it a unique technological resource within the EU research landscape.
The work involved reverse-engineering earlier prototypes, designing improved components compatible with multiple spectrometer systems, and integrating advanced control software. The resulting instrument is mobile, robust, and “vendor-agnostic”, meaning it can operate across different NMR platforms — a feature that greatly enhances its accessibility for future users.
Alongside the hardware development, the we developed new software tools for experiment control, data acquisition, and analysis. Novel methods for non-uniform data sampling and signal processing were implemented to enable faster, more sensitive measurements of protein dynamics.
Although biochemical applications were delayed by the time required to build and validate the new instrument, significant groundwork was laid for upcoming studies on model proteins such as γD-crystallin and IAPP, which are linked to cataract formation and type II diabetes, respectively.
The project also established essential laboratory infrastructure for future independent research, including protein expression and purification facilities, electronic workshops, and computational environments for reproducible data analysis. Training activities in programming, instrument design, and scientific communication helped to deliver Europe's first PJ-NMR instrument. Together, completion of this project established Europe’s position in this emerging field.
The work involved reverse-engineering earlier prototypes, designing improved components compatible with multiple spectrometer systems, and integrating advanced control software. The resulting instrument is mobile, robust, and “vendor-agnostic”, meaning it can operate across different NMR platforms — a feature that greatly enhances its accessibility for future users.
Alongside the hardware development, the we developed new software tools for experiment control, data acquisition, and analysis. Novel methods for non-uniform data sampling and signal processing were implemented to enable faster, more sensitive measurements of protein dynamics.
Although biochemical applications were delayed by the time required to build and validate the new instrument, significant groundwork was laid for upcoming studies on model proteins such as γD-crystallin and IAPP, which are linked to cataract formation and type II diabetes, respectively.
The project also established essential laboratory infrastructure for future independent research, including protein expression and purification facilities, electronic workshops, and computational environments for reproducible data analysis. Training activities in programming, instrument design, and scientific communication helped to deliver Europe's first PJ-NMR instrument. Together, completion of this project established Europe’s position in this emerging field.
Pressure-Jump NMR introduces a new experimental paradigm that bridges a critical gap in molecular science: the ability to control and monitor fast, transient processes under physiological conditions. Traditional NMR and other structural techniques capture molecules in equilibrium; PJ-NMR makes it possible to observe how they change and interact in real time.
This breakthrough has far-reaching implications:
- Scientific impact: PJ-NMR enables the direct study of protein folding, aggregation, and phase separation kinetics — phenomena at the heart of many biological and pathological processes. By providing atom-level insight into how aggregates and liquid droplets form, the method can reshape our understanding of diseases such as Alzheimer’s and diabetes.
- Technological impact: Establishing Europe’s first operational PJ-NMR platform opens new possibilities for collaboration between academic and industrial researchers, particularly in biotechnology and pharmaceutical development. The technology’s modular, open-source design allows it to be reproduced and improved by other laboratories, accelerating innovation across the continent.
- Capacity building: The project strengthened European infrastructure for advanced NMR research by combining hardware innovation, open-source software development, and reproducible data practices. It also supported the researcher’s transition to independence through the establishment of a new laboratory and a Swedish Research Council Starting Grant.
In the medium term, PJ-NMR is expected to contribute to the discovery of molecular mechanisms that can inform new therapeutic strategies or screening methods for aggregation inhibitors. Over the longer term, it may support the design of biomolecules and materials that exploit controlled phase separation — an area with growing relevance in biotechnology and medicine.
This breakthrough has far-reaching implications:
- Scientific impact: PJ-NMR enables the direct study of protein folding, aggregation, and phase separation kinetics — phenomena at the heart of many biological and pathological processes. By providing atom-level insight into how aggregates and liquid droplets form, the method can reshape our understanding of diseases such as Alzheimer’s and diabetes.
- Technological impact: Establishing Europe’s first operational PJ-NMR platform opens new possibilities for collaboration between academic and industrial researchers, particularly in biotechnology and pharmaceutical development. The technology’s modular, open-source design allows it to be reproduced and improved by other laboratories, accelerating innovation across the continent.
- Capacity building: The project strengthened European infrastructure for advanced NMR research by combining hardware innovation, open-source software development, and reproducible data practices. It also supported the researcher’s transition to independence through the establishment of a new laboratory and a Swedish Research Council Starting Grant.
In the medium term, PJ-NMR is expected to contribute to the discovery of molecular mechanisms that can inform new therapeutic strategies or screening methods for aggregation inhibitors. Over the longer term, it may support the design of biomolecules and materials that exploit controlled phase separation — an area with growing relevance in biotechnology and medicine.