Biophysical approaches in a structure-guided SMYD3 ligand discovery

dc.contributor.authorTalu, Martin Johannes
dc.contributor.departmentChalmers tekniska högskola / Institutionen för biologi och biotekniksv
dc.contributor.examinerNorbeck, Joakim
dc.contributor.supervisorTalibov, Vladimir O.
dc.date.accessioned2020-02-11T10:04:28Z
dc.date.available2020-02-11T10:04:28Z
dc.date.issued2019sv
dc.date.submitted2019
dc.description.abstractIn eukaryotic cells, DNA is wrapped around nucleosomal cores formed of protein heterooctamers, which consist of core histones. Nucleosomes are the main units of chromatin organization. Chromatin exist in two states - either as eu- or heterochromatin, which either promotes or silences gene expression as a consequence of its packing states. Eukaryotic cells have developed epigenetic regulation to control the chromatin state, and to guarantee a high level of differentiation. The basis of epigenetic regulation are patterns of post-translational modifications of the nucleosomal proteins. These modifications are performed by epigenetic enzymes. This thesis focuses on one of these enzymes - the human lysine methyltransferase SMYD3. SMYD3 is also capable of interacting with certain cytosolic proteins, such as the molecular chaperone HSP90 - the human Heat Shock Protein 90. Both proteins are of high interest in the drug research and development landscape, as a drastic change in their activity and expression levels have both been shown to be related to several cancers or neurodevelopmental diseases. In this work, various truncated forms of HSP90 were produced and probed for their interactions with SMYD3 using surface plasmon resonance-based biosensor technology. A previously reported interaction of SMYD3 with the C-terminal domain of HSP90 was confirmed, with an affinity discovered to be KD = 1.3 × 10−5M. Additionally, the biosensor-based assay was used to test potential ligands of SMYD3, including low affinity fragment-like organic molecules. To complement the study, extensive crystallization and co-crystallization trials were carried out with SMYD3. As a result, conditions for the formation of various crystal forms of SMYD3 were mapped, with the best crystal form found to have high stability and good diffraction properties. A set of experiments presented herein develops expertise in the tools one can use for an efficient and rational ligand discovery campaign targeting SMYD3 histone methyltransferase.sv
dc.identifier.coursecodeBBTX60sv
dc.identifier.urihttps://hdl.handle.net/20.500.12380/300679
dc.language.isoengsv
dc.setspec.uppsokLifeEarthScience
dc.subjectSMYD3sv
dc.subjectHSP90sv
dc.subjectSPRsv
dc.subjectXRDsv
dc.subjectMSTsv
dc.subjectTSAsv
dc.subjectdrug discoverysv
dc.titleBiophysical approaches in a structure-guided SMYD3 ligand discoverysv
dc.type.degreeExamensarbete för masterexamensv
dc.type.uppsokH
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Biophysical approaches in a structureguided SMYD3 ligand discovery Master’s thesis in Biotechnology MARTIN JOHANNES TALU
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