FACULTY
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Mark E. Bowen Website: https://www.bowenlab.org/ |
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Research |
The Bowen Lab focuses on the membrane-associated guanylate kinase (MAGuK) family of scaffold proteins, which are key organizers of excitatory neurotransmission and are implicated in diseases such as neurodegeneration following stroke, autism, epilepsy and schizophrenia. Scaffolds determine the outcome of signal transduction by controlling the location of receptors and connecting them to downstream effectors. Studying scaffolds in vivo has been hampered by the complexity and redundancy of multiple isoforms present in the brain. Our approach is to reconstitute these cellular complexes on biological membranes to uncover how protein structure and posttranslational modifications, such as phosphorylation and palmitoylation, shape their scaffolding activity. By taking a reductionist approach, we can learn the rules that govern the spontaneous assembly of proteins within the postsynaptic density of excitatory neurons. The proteins we study contain intrinsic disorder meaning they lack the fixed structures that give rise to activity in enzymes yet they mediate critical synaptic functions. The lack of a fixed structure challenges conventional approaches to structural biology so we use fluorescence microscopy to follow their transient interactions and refine molecular models for their dynamic structures. Harnessing imaging methodology for protein structure determination allows us to study previously intractable complexes. Our primary approach is to image at the level of single molecules, which allows us to follow the composition, structure and dynamics of an individual scaffold and its complexes with synaptic proteins. By combining the latest in single molecule methods with our working reconstitution of the postsynaptic membrane, we are working clarify the dynamic assembly of proteins that govern the response to glutamate neurotransmitters at the excitatory synapse.
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Selected |
Yanez-Orozco, I.S., Levesque B., Ma, J., Wang, B., Spencer, M., Adariani, S.R., Ding, F.*, Bowen, M. E.*, Sanabria, H*. “Identifying Weak Interdomain Interactions that Stabilize the Supertertiary Structure of the N-Terminal Tandem PDZ Domains of PSD-95” Nature Communications (2018) in press (* co-corresponding authors) Hellenkamp, B., Schmid, S., Doroshenko, O., Opanasyuk, O., Kühnemuth, R., Adariani, S.R., Barth, A., Birkedal, V., Bowen, M. E., Chen, H., Cordes, T., Eilert, T., Fijen, C., Götz, M., Gouridis, G., Gratton, E., Ha, T., Hanke, C.A., Hartmann, A., Hendrix, J., Hildebrandt, L.L., Hohlbein, J., Hübner, C., G., Kallis, E., Kapanidis, A.N., Kim, J., Krainer, G., Lamb, D. C., Lee, N.K., Lemke, E.A., Levesque B., Levitus, M, McCann, J.J., Naredi-Rainer, N., Daniel Nettels, Ngo, T., Qiu, R., Röcker, C., Sanabria, H., Schlierf, M, Schuler B., Seidel, H., Streit, L., Tinnefeld, P., Tyagi, S., Vandenberk, N., Weninger, K.R., Wünsch B., Yanez-Orozco, I.S., Michaelis, J., Seidel, C.A.M., Craggs, T.D., Hugel, T. “Precision and accuracy of single-molecule FRET measurements - a worldwide benchmark study.” (2017) arXiv arXiv:1710.03807v2 Pedersen S.W., Albertsen L, Moran G.E., Levesque B., Pedersen S.B., Bartels L., Wapenaar H., Yei F., Zhang M., Bowen M.E., Strømgaard K. “Site-specific phosphorylation of PSD-95 PDZ domains reveals fine-tuned regulation of protein-protein interactions.” ACS Chem Biol. (2017) Sep 15; 12(9):2313-2323 (PMID: 28692247) Cabail M.Z., Li S., Lemmon E., Bowen M.E., Hubbard S.R., Miller W.T. "The insulin and IGF1 receptor kinase domains are functional dimers in the activated state." Nature Communications (2015) Mar 11; 6:6406. (PMID: 25758790) McCann J.J., Choi U.B., Bowen M.E. "Reconstitution of Multivalent PDZ Domain Binding to the Scaffold Protein PSD-95 Reveals Ternary-Complex Specificity of Combinatorial Inhibition." Structure (2014) Oct 7; 22(10):1458–1466. (PMID: 25220472) Choi U.B., Kazi R., Stenzoski N., Wollmuth L.P., Uversky V.N., Bowen M.E. “Modulating the Intrinsic Disorder in the Cytoplasmic Domain Alters the Biological Activity of the N-Methyl-D-aspartate-sensitive Glutamate Receptor.” Journal of Biological Chemistry. (2013) Aug 2; 288(31):22506-15. (PMCID: PMC3829338). McCann J., Zheng, L., Rohrbeck, D., Felekyan, S., Kühnemuth, R., Sutton, R.B., Seidel, C.A.M., Bowen, M.E. “Supertertiary Structure of the Synaptic MAGuK Scaffold Proteins is Conserved” Proceedings of the National Academy of Sciences (2012) Sep 25;109(39):15775-80. (PMCID: PMC3465453). Choi, U.B., Xiao, S., Zheng, L., Bowen, M.E. “Effect of Src Phosphorylation on the Disordered C-Terminal Domain of the NMDA Receptor Subunit GluN2B.” Journal of Biological Chemistry (2011) Aug. 26 286:29904-29912 (PMCID: PMC3191031). McCann J., Zheng, L., Chiantia, S., Bowen, M.E. “Domain Orientation in the Tandem PDZ Supramodule from PSD-95 is Maintained in the Full-Length Protein” Structure (2011) 19, 810-820. (PMCID: PMC3116789). Choi, U.B., McCann J., Weninger, K., Bowen, M.E. “Beyond the random coil: stochastic conformational switching in intrinsically disordered proteins.” Structure (2011) Apr 13; 19(4):566-76. (PMCID: PMC3075556).
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