Old Road Campus Research Building
Dr, Principal Investigator, Group Head
The Biotechnology group at the SGC in Oxford is primarily responsible for generating the pipeline of clones and determining which proteins are expressed in a soluble and stable form suitable for structural and functional studies. We have developed and optimised protocols for high-throughput ligation independent cloning, test expression and purification, large scale expression, protein production and characterisation by mass spectrometry. The target list stems from protein areas of interest to the Biology PIs within the SGC and includes epigenetics, kinases, organelle biogenesis, growth factor signalling, DNA-RNA processing and IMPs. The support and responsiveness we provide to the epigenetics and the soluble and IMP structural projects, helps to advance the knowledge in these fields and our rate of success.
The platform established by the Biotechnology team, has enabled the site at Oxford to generate 400 novel human protein structures and 2 integral membrane protein structures to date. The group collaborates and interacts closely with the other SGC teams, to develop methods for increasing protein expression, parallel processing and driving output. To date, we have been using both E. coli and baculovirus/insect cell expression systems for protein production. However initiating new method development projects is an important aspect of our role within the organisation to ensure we find ways of producing the most challenging proteins on the target list, many of which are also of high value to the scientific community. Consequently, we are currently setting up platforms for producing proteins in Saccharomyces cerevisiae and mammalian cells, including BacMam technology. We may also explore other expression systems in the future. In addition to enhancing protein expression levels, we also aim to reduce our cost per structure by seeking alternative cheaper reagents, consumables and increasing throughput further by miniaturising the processes and increasing automation where possible. Discovering ways to successfully produce the most challenging human proteins would open many doors to our long-term goals as an organisation of understanding how proteins function independently, with partners and also how they interact with drugs.
A lower X-gate in TASK channels traps inhibitors within the vestibule
Rodstrom KEJ. et al, (2020), NATURE
The structural basis of lipid scrambling and inactivation in the endoplasmic reticulum scramblase TMEM16K
Bushell SR. et al, (2019), Nature Communications, 10
A genetics-led approach defines the drug target landscape of 30 immune-related traits
Fang H. et al, (2019), Nature Genetics, 51, 1082 - 1091
High-Throughput Site-Directed Mutagenesis.
Strain-Damerell C. and Burgess-Brown NA., (2019), Methods Mol Biol, 2025, 281 - 296
Structures of DPAGT1 Explain Glycosylation Disease Mechanisms and Advance TB Antibiotic Design.
Dong YY. et al, (2018), Cell, 175, 1045 - 1058.e16