Uncovering Molecular Interactors of Ca2+/Calmodulin-Dependent Protein Kinase II (CaMKII)

Presenter

Brooke Mary Abouhamad

Campus

UMass Amherst

Sponsor

Margaret Stratton, Department of Biochemistry and Molecular Biology, UMass Amherst

Schedule

Session 3, 1:30 PM - 2:15 PM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A48, Campus Center Auditorium, Row 3 (A41-A60) [Poster Location Map]

Abstract

Ca2+/calmodulin-dependent protein kinase II (CaMKII) is a crucial protein in the biological process of memory formation. CaMKII is a serine/threonine kinase with unique characteristics and is required for long-term potentiation (LTP), the cellular basis of learning and memory. Being the most concentrated enzyme at the postsynaptic density, CaMKII is surrounded by numerous proteins including receptors, scaffold proteins, enzymes, and cytoskeletal proteins. Currently, we know that CaMKII interacts directly with glutamate receptors (such as NMDAR), Tiam1, alpha-actinin, F-actin, etc (Özden et al., 2022), all aiding in its function. Although some interactors are known, there are likely other interactors. This research focuses on determining all proteins that interact with CaMKII in hippocampal neurons. To test this, CaMKII was used as a protein bait for proximity labeling in Human Endothelial Kidney (HEK) cells and hippocampal neurons. A mutant biotin ligase, miniTurbo, is fused onto the N-terminus of CaMKII to allow for this proximity labeling to occur. Streptavidin pulldowns of HEK and neuronal lysates were performed, processed by western blotting, and mass spectroscopy to confirm proteomic constituents. With these experiments, it is expected to find that CaMKII interacts with NMDARs, densin, alpha-actinin, due to their high abundance at the postsynaptic density. In the future, we will perform proteomics on all CaMKII paralogs, to find novel interacting partners and their significance in function. These techniques will allow us to identify molecular targets and drive our understanding of CaMKII function and regulation in cells.

Keywords

Memory , Protein Interactors, proximity labelling , CaMKII, Long term potentiation

Research Area

Biological Organisms

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