Titin has a repetitive beads-on-a-string arrangement of small domains. Many of these domains are structurally similar. Therefore, information from one domain can be used to predict how another related domain would behave should it have a SNP. In this work I have firstly identified that this assumption – that information form one can be used to predict another’s behaviour- is correct and then used it to expand the knowledge that we have from well-characterised areas to those areas that have not been studied directly. Training in mSNP annotation was received at Prof Richard’s lab (Généthon, Évry France), During the training exchange I was made aware of several publicly available databases and how to correctly interpret the information with in them. In particular, a data set of healthy individuals over 60 from Brazil (of mixed ethnic backgrounds) was used to identify tolerated SNVs.
During this work a total 10 new 3D domain structures across four areas of titin have been solved. increasing the structural coverage of titin domains, allowing for the better assessment of uniqueness and repetition across the titin chain. These will be deposited in the PDB - a public database of structures- when their accompanying papers detailing their biological significance are also published. The preparation of samples for crystal structure experiments resulted in the development of a new phosphorylation purification method (Adams, M et. al., 2019, Protein Journal. 38(2):181-189). In addition, due to the expertise gained from titin domain structural analysis, I also contributed towards a biomimetics project resulting in two publications (Hill, CJ et. al. Advanced Materials. 31(17):e1807521, 2019; Nesterenko, Y et. al., 2018, International Journal of Molecular Sciences. 20(17), 4299).
Several scripts have been developed to extract, organise and assimilate the data into diagnostic scores. Upon publication of the final tool, these will be made publicly available for those wishing to use or alter them. During this work to determine which methods would be suitable for incorporation into the molecular diagnostics pipeline, I undertook a series of detailed analysis to serve as case studies for structural mSNP assessment in titin. Firstly, I expanded on previous work using molecular dynamics to assess the effect of mSNPs on interdomain conformations (Fleming, JR et. al., 2020, Journal of Biomolecular Structure and Dynamics). Then, I investigated a cardiomyopathy-linked mSNP involved in a stretch induced molecular phenotype (manuscript submitted for review), as well as the effects of SNPs on a unique area of titin involved in binding stress response factors (manuscript in preparation). Details of additional methods and supporting data, confirming the applicability the chosen approaches integrated into the final tool and creation of the overall pipeline, are currently being prepared into several manuscripts. The final tool will be released and disseminated to the clinical community upon publication of these supporting manuscripts.
To support the dissemination of my work to a general audience I partook in several outreach activities. In 2019 I participated in two events associated with the European Researchers’ Night. The first was giving a public engagement talk on the project “Wrestling with the muscular giant, Titin” in Ely, UK. The second I participated in the Cambridge Life Lab in Peterborough, which was publicised in the local press. This was a series of hands on experiments for children set up in a shopping centre. This event was very popular (with over 300 visits in the day) and provided the opportunity not only engage children and also their parents. I have also created a profile as part of the women in science (WISE) campaign role model initiative (
https://www.myskillsmylife.org.uk/my-skills-role-model/1002671(odnośnik otworzy się w nowym oknie)) to help inspire young girls into STEM careers. At a local level, I have become a member of the nascent University of Konstanz 500 women scientist ‘pod’ (
https://500womenscientists.org/(odnośnik otworzy się w nowym oknie)). We aim to increase local awareness of issue affecting female researchers and contribute to global schemes to advocate for science in society and women in science (such as organising wiki-thons where female academics profiles are written and translated).
