[ Research Interests ] [ Representative Publications ]

Michael Edidin Professor Department of Biology B.S. University of Chicago Ph.D. University College London Postdoctoral The Weizmann Institute Harvard Medical School

Department of Biology Johns Hopkins University 3400 North Charles Street Baltimore, MD 21218-2685 U.S.A. Office Telephone: Lab Telephone: Department Fax: Email: 410.516.7294 410.516.7295 410.516.5213 edidin@jhu.edu Office- Mudd 38A Lab- Mudd 45

Research Interests Cell membranes are complex two-dimensional arrays of mobile, interacting molecules. My laboratory uses cell biology, biophysics, especially fluorescence methods, biochemistry and immunology to study membrane dynamics and organization in cells ranging from lymphocytes to epithelial cells. All of our work on membranes arises from interests in transplantation immunology, especially in the cell biology of class I MHC molecules. We aim to understand: 1) the intracellular traffic of class I MHC molecules during and after peptide loading, 2) the relationship between plasma membrane biophysics and antigen presentation by class I molecules and 3) the way in which viral proteins interfere with both these processes. Our methods of analysis concentrate on two microscope-based physical techniques, fluorescence recovery after photobleaching, FRAP, to measure lateral diffusion, and fluorescence resonance energy transfer, FRET, to measure molecular proximity and clustering. Work with these techniques as well as with other advanced microscopies, including deconvolution microscopy and total internal reflection microscopy, is done in a strong Departmental imaging facility with ample hardware and technical support. Using genetically fluorescent class I  MHC molecules (tagged with GFP and its derivatives) we have dissected the organization of these molecules and their specialized chaperones in the endoplasmic reticulum, ER. Thus far we know that nascent class I molecules remain in the ER after peptide loading and that they interact with a number of factors some of which may carriers for export from the ER. We also have some evidence that the ER membrane is organized into domains with specialized function. Experiments are planned to study ER domain organization in detail and to resolve the nature of class I-associated molecules regulating their ER export. The same reagents and methods can be used to understand the ways in which viral proteins that suppress expression interact with class I molecules in the ER. We also investigate the way in which antigen presentation to T cells and NK cells is affected changes in organization of class I MHC molecules at the cells surface. We are concentrating on techniques that vary the mobility and of the class I molecules and on methods that change the extent to which they are clustered. It appears that cholesterol depletion of plasma membranes has large effects on antigen presentation; these effects can be mimicked by drugs that affect the membrane skeleton and by class I MHC molecules that have been engineered so that they can be crosslinked and clustered by small membrane-permeable compounds. We have shown that increasing the size of class I MHC clusters enhances presentation of low concentrations of antigens. We have some evidence that changes in class I MHC clustering occur naturally when cells are activated for antigen presentation. It is thought that membrane proteins and lipids are not randomly distributed in the fluid lipid bilayer. Rather, both proteins and lipids may associate to form domains. Most of the evidence for these domains is indirect. Many domains are smaller than the resolution of the light microscope. We have built a "super-resolution" light microscope, a near-field scanning optical microscope, or NSOM, which images cells labeled with fluorescent antibodies, but with a resolution of better than 50nm. Using this microscope we plan to image the patchiness of surfaces of living cells, and to follow changes in this patchiness when cells present antigen. A particular type of lipid domain, a 'lipid raft', has been suggested as important for trafficking of some membrane lipids and proteins, and for the assembly of signaling complexes after surface receptors bind their ligands. We are probing for these rafts using fluorescence resonance energy transfer. Our fluorescent probes may be either labeled antibodies, or variants of GFP. Representative Publications Everett, M.W. and Edidin, M. 2007. Tapasin Increases Efficiency of MHC I Assembly in the Endoplasmic Reticulum but Does Not Affect MHC I Stability at the Cell Surface. The Journal of Immunology. 179:7646-52. Fooksman, D.R., Edidin, M., and Barisas, B.G. 2007. Measuring rotational diffusion of MHC class I on live cells by polarized FPR. Biophys Chem. 130:10-6. Shaikh SR, Edidin MA. 2006. Membranes are not just rafts. Chem Phys Lipids.144(1):1-3. Ladasky, J.J., Boyle, S., Seth, M., Li, H., Pentcheva, T., Abe, F., Steinberg, S.J., Edidin, M. 2006. Bap31 enhances the ER export and quality control of human class I MHC molecules. J. Immuol. 177:6172-81. Fooksman DR, Gronvall GK, Tang Q, Edidin M. (2006). Clustering class I MHC modulates sensitivity of T cell recognition. J, Immunol. 176(11):6673-80. Capps, G.G, Pine, S., Edidin, M., and Zúñiga, M.C. (2004) Short Class I MHC Cytoplasmic Tails Differing in Charge Detect Arbiters of Lateral Diffusion in the Plasma Membrane Biophys. J. 86, 896-909. Kwik, J., Boyle, S., Fooksman, D. Margolis,L., Sheets, M.P. and Edidin, M. (2003) Membrane cholesterol, Lateral mobility & the PI(4,5)P2-dependent organization of cell actin. Proc. Natl. Acad. Sci. USA 100, 13964-13969. Rocheleau, J.V., Edidin, M. and Piston, D.W. (2003) Intrasequence GFP in class I MHC molecules, a rigid probe for fluorescence anisotropy measurements of the membrane environment. Biophys. J. 84, 4078-4086. Tang Q. and Edidin, M. (2003) Lowering the barriers to random walks on the cell surface. Biophys. J. 84(1) 400-407. Edidin, M. (2003) The State of Lipid Rafts: from Model Membranes to Cells. Annu. Rev. Biophys. Biomolec. Struct. 32, 257-283. Spiliotis, E. T., Pentcheva, T. and Edidin, M. (2002) Probing for membrane domains in the endoplasmic reticulum; retention and degradation of unassembled MHC class I molecules. Mol. Biol. Cell 13, 1566-1582. Pentcheva, T., Spiliotis, E.T. and Edidin, M. (2002) Tapasin is retained in the endoplasmic reticulum by dynamic clustering and exclusion from endoplasmic reticulum exit sites. J. Imunol. 168, 1538-1541. Fahmy, T., J.G. Bieler, M. Edidin and J. Schneck. (2001).Increased TCR avidity after T cell activation: A mechanism for sensing low-density antigen. Immunity 14, 135-143. Spiliotis, E.T., H. Manley, M. Osorio, M. C. Zúñiga, and M. Edidin. (2000) Selective Export of MHC Class I Molecules from the ER after Their Dissociation from TAP. Immunity 13, 841-851.

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