En singelmolekyl-fälla av DNA origami för att studera självskapade värmechocker hos enzym

Tidsperiod: 2017-07-01 till 2020-06-30

Projektledare: Alan Shaw

Finansiär: Vetenskapsrådet

Bidragstyp: Bidrag för anställning eller stipendier

Budget: 3 150 000 SEK

In this proposal, we aim to explore the uncharted interactions between the heat released from an exothermic enzymatic reaction, and the enzyme’s structure and function. To achieve this goal, we chose pyruvate kinase (PK) as the model enzyme, as the heat released by its catalytic reaction is easily tunable. We will integrate my skills in DNA origami and the host laboratories’ excellent track record in optical tweezers and single molecule FRET to develop a novel single molecule trap that can simultaneously probe the functional response of PK to its heat of reaction.In the first year we will perform molecular cloning of PK and develop the DNA origami single molecule trap. Study of PK’s response to its heat of reaction will be conducted in the second and part of the third year. The remaining time will be invested on the communication of results.The work in this project represents a milestone in our understanding of heat flow in biological systems, and will break the long-standing belief that the heat generated from exothermic enzymatic reactions does not affect the enzyme itself. Understanding the mechanistic interactions between the heat release by PK and PK itself will open up new possibilities in cancer therapy: development of drugs that target this process, tuning PK activity to impede cancer growth.The outstanding scientific milieu in the host laboratories will grant me a unique set of skills that will have a momentous impact in my future career as an independent researcher.