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Informationen zum Autor Dr. Natalia Jura is an Associate Professor and an Investigator in the Department of Cellular and Molecular Pharmacology at the Cardiovascular Research Institute within the School of Medicine at the University of California, San Francisco (UCSF). She is also an Associate Director of the Quantitative Biosciences Institute at UCSF. Dr. Jura's group at UCSF focuses on understanding how soluble protein kinases and membraneassociated receptor kinases assemble into functional complexes and regulate their signaling through molecular interactions with regulatory proteins. Her group also investigates alternative non-catalytic roles of protein kinases as scaffolds in cellular signaling pathways. They apply this knowledge for design of small molecule inhibitors that target these poorly understood kinase functions in human diseases. Dr. Jura received her M.S in biochemistry from Jagiellonian University in Krakow, Poland and her Ph.D. in molecular and cellular biology from Stony Brook University. James Murphy is Associate Professor and the head of the Inflammation Division at the WEHI (formerly known as The Walter and Eliza Hall Institute of Medical Research) in Melbourne and is closely associated with The University of Melbourne and The Royal Melbourne Hospital. James’ lab studies the protein-protein interactions that underpin signal transduction. Much of his work is focused on understanding the molecular mechanisms by which protein kinases and their relatives, pseudokinases, regulate cell signaling. Their work on the pseudokinase, MLKL, provided a template for developing a detailed understanding of how the remaining ~50 uncharacterized pseudokinases modulate cell signaling.
List of contents
1. Production and Purification of the PEAK pseudokinases for structural and functional studies
2. Structural biology and biophysical characterisation of Tribbles pseudokinases
3. Detecting endogenous TRIB protein expression and its downstream signalling
4. Analysis of human Tribbles 2 pseudokinase
5. Expression, purification and examination of ligand-binding to IRAK pseudokinases
6. Characterization of pseudokinase ILK-mediated actin assembly
7. Biochemical examination of Titin pseudokinase
8. Approaches to study pseudokinase conformations
9. CRISPR editing cell lines for reconstitution studies of pseudokinase function
10. Proximity ligation approaches to define the interactomes of pseudokinases
11. Determining the role of the pseudokinase domain in guanylyl cyclase A and B signaling
12. Strategies to study and stimulate bacterial pseudokinases
13. Dynamics of Kinases and Pseudokinases by HDX-MS
14. In-cell thermal shift assay for drug binding analysis to receptor pseudokinases
15. Pseudoenzymology of Kinase Suppressor of Ras
16. Computational and evolutionary insights into pseudokinase mechanisms
17. Methods to identify small molecule allosteric modulators of the STRAD pseudokinase
18. Pharmacological targeting of the pseudokinase HER3
19. Considerations and analysis of the unusual pseudokinase BUBR1
20. The pseudokinase domain of receptor guanylyl cyclases: binding of ATP and allosteric regulation of guanylyl cyclase activity
21. Methods for discovering catalytic activities for pseudokinases
22. Efficient Expression, Purification, and Visualization by Cryo-EM of Fully-Glycosylated Unliganded HER3
23. An effective strategy for ligand-mediated pulldown of the HER2/HER3 receptor complex and cryo-EM structure determination at low sample concentrations
24. Identification of small molecule binders for pseudokinases
25. Identification and characterization of Tyk2-JH2 pseudokinase domain inhibitors
26. Monitoring the ATPase activity of Bud32 and its role in tRNA binding
