Our Methods
Recombinant Protein Production
We produce recombinant proteins using various host systems, including bacteria, yeast, insect, and mammalian cells. This diverse approach allows us to tailor protein expression to the specific requirements of each research project. We have developed a custom expression system to facilitate switching between expression systems (Altmannova et al., Protein Science 2021).
We generate a wide range of proteins, from small fragments to entire complexes, enabling detailed studies of their biological functions. We employ advanced ÄKTA FPLC chromatography systems, ensuring high-quality purification suitable for structural and functional analyses.
We generate a wide range of proteins, from small fragments to entire complexes, enabling detailed studies of their biological functions. We employ advanced ÄKTA FPLC chromatography systems, ensuring high-quality purification suitable for structural and functional analyses.
In vitro reconstituion
Protein complexes underpin virtually all cellular functions, with many essential processes, such as meiotic recombination, relying heavily on interactions between proteins and nucleic acids. Our laboratory focuses on reconstructing these large, intricate complexes, particularly those involving nucleosomes, to elucidate the underlying principles governing their assembly and function. This approach provides critical insights into the fundamentals of meiosis.
Biophysical Characterisation
Our laboratory employs advanced biophysical techniques, including mass photometry and SEC-MALS, to accurately determine the composition and stoichiometry of protein complexes. We utilise ITC, SPR, and MST to measure precise binding affinities within macromolecular interactions. These methods collectively enable us to build detailed mechanistic models of how protein complexes facilitate critical cellular processes.
Hybrid Structural Biology
We integrate computational modelling with experimental data from chemical cross-linking mass spectrometry (XL-MS), small-angle X-ray scattering (SAXS), and single-particle cryo-electron microscopy (cryo-EM). This hybrid approach allows us to construct comprehensive structural models of protein and nucleic acid complexes, revealing insights into their architecture and function. Such insights allow us to create carefully designed mutants to study in vivo.
Yeast Genetics
Meiosis is a fundamental and evolutionarily conserved biological process. Budding yeast serves as an ideal model organism, offering a highly tractable genetic system for investigating the molecular machinery underpinning meiotic recombination. By introducing specific mutations, we can examine their impact on recombination initiation, crossover formation, and spore viability, providing deep insights into the basic principles of meiosis.
