In this month’s lab profile we highlight the work of Dr Ravi Nookala, a structural biologist working in the lab of Professor Sir Tom Blundell in the Department of Biochemistry at the University of Cambridge.
Dr Nookala is working on the structural biology of proteins involved in BHD syndrome. Knowing the structures of these proteins, for instance identifying conserved motifs or folds, would greatly assist in understanding their function. Not only that, structural information is also invaluable for developing drugs and therapies. For example, by finding a potential binding site on a protein, molecules can be tailor made to fit to this site and therefore bind with high affinity. Binding of a drug with high affinity to a protein can alter the protein’s behaviour, for example by modifying its activity, turnover or post-translational modification.
X-ray crystallography is a technique commonly used to determine the structure of proteins. The first step in this technique is to produce a large amount of recombinant pure protein by expression in systems such as bacteria. The protein is then put into crystallisation trials where a range of conditions are tested to promote the formation of protein crystals. Once crystals have grown, they are frozen and mounted on an X-ray detector. X-rays are fired at the crystal and the diffraction image is collected. After applying several mathematical formulae to this data, a 3D image of the protein is generated.
Due to disordered regions within a protein, whole proteins are often difficult to crystallise. In this case, the protein can be split up into domains and the disordered regions removed, which can aid the crystallisation process. This is exactly what Dr Nookala, research assistant Beata Blaszczyk and PhD student Angela Pacitto are doing with FLCN. Dr Nookala has successfully determined the structure of the C-terminal domain of FLCN and work is now on-going to understand the rest of the FLCN protein and to characterise its interaction with other proteins.
The following publication highlights Dr Nookala’s previous work on Fanconi Anaemia and gives further examples of the type of structural work that is performed in this lab:
- Nookala RK, Hussain S, & Pellegrini L (2007). Insights into Fanconi Anaemia from the structure of human FANCE. Nucleic acids research, 35 (5), 1638-48 PMID: 17308347