Revolutionising macromolecular crystal structure determination through routine multi-dataset analysis

Samenvatting

Crystallography has been the workhorse of protein structure determination for 60 years and remains the premier method for obtaining high-resolution structures in the search for new therapeutics. During that time, the principles for constructing protein models – human-centred visual interpretation of a single electron density map – have remained largely unchanged. Fundamental ambiguity in the experimental data (from averaging over the population of conformations in the crystal) makes modelling regions with multiple conformations insensitive and imprecise, and makes reliable modelling intractable even for automated methods. Moreover, information regarding protein dynamics is lost.
To overcome the ambiguity in crystallographic data, I am currently developing methods to model series of crystallographic datasets simultaneously, where the population of conformations is slightly perturbed from crystal to crystal. These “perturbation series” remove the ambiguity in crystallographic data, and thus lead to more accurate, reliable and detailed crystallographic models without relying on human interpretation. Furthermore, as a natural result of these perturbation series, we can extract correlated changes in protein structures at the atomic level, enabling identification of subtle structural effects such as allostery.
I am developing these methods for medium-throughput crystallographic fragment screening (100-1000s of datasets). In this proposed project, I will develop experimental protocols to allow multi-dataset methods to be applied “routinely” by all protein crystallographers (using approximately 10 datasets). I will demonstrate the benefit of routine multi-dataset protocols by applying them to a wide variety of biologically-relevant systems with collaborators in the Netherlands and internationally. These applications will demonstrate the effectiveness of multi-dataset methods on “everyday” crystallographic experiments and revolutionise the way that protein structures are determined. Given the scope and potential audience for these techniques, this project is positioned to have a large impact on the way that scientists determine and use protein structures in the Netherlands and all over the world.

Kenmerken

Projectnummer

VI.Veni.192.143

Hoofdaanvrager

Dr. N.M. Pearce

Verbonden aan

Universiteit Utrecht, Faculteit Bètawetenschappen, Departement Scheikunde

Looptijd

01/03/2020 tot 28/02/2023