Abstract
Human interferon-gamma (hIFN\(\gamma \)) is a key immunomodulating secretory glycoprotein. Although glycosylation is not necessary for its activity, it does affect the physico-chemical properties of the cytokine. Recently, a technology was developed for the secretory expression of glycosylated hIFN\(\gamma \) and its highly prone to aggregation mutant K88Q in insect cells. In addition, the proteins were labeled with specific tag peptides added to their N-termini. It was experimentally observed, that the obtained fusion proteins have significantly reduced biological activity and when glycosylated, they were resistent to enterokinase action and the tag could not be removed. Here we report the development of in silico models of glycosylated His\(_6\)-FLAG-hIFN\(\gamma \) fusion proteins and employ long-scale molecular dynamics (MD) simulations to explain these unexpected experimental results and to study in detail the effect of the tags and glycosylation on the structure and dynamics of the fusion glycoproteins.
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Acknowledgements
This work was supported in part under the Programme for young scientists’ career development at the Bulgarian Academy of Sciences (DFNP-17-146/2017) and under Grants DN-11/20/2017 and DNTS-Austia-01-2/2013 of the Bulgarian Science Fund. Computational resources were provided by the supercomputer Avitohol@BAS and the HPC Cluster at the Faculty of Physics of Sofia University “St. Kl. Ohridski”.
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Lilkova, E., Ilieva, N., Petkov, P., Krachmarova, E., Nacheva, G., Litov, L. (2021). Molecular Dynamics Simulations of His\(_6\)-FLAG-hIFN\(\gamma \) Fusion Glycoproteins. In: Georgiev, I., Kostadinov, H., Lilkova, E. (eds) Advanced Computing in Industrial Mathematics. BGSIAM 2018. Studies in Computational Intelligence, vol 961. Springer, Cham. https://doi.org/10.1007/978-3-030-71616-5_23
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DOI: https://doi.org/10.1007/978-3-030-71616-5_23
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