The objectives of this proposal are (1) to extend current information about immunoglobulin (Ig) structure, derived from X-ray crystallographic studies of IgG1 antibodies, to the full range of Ig isotypes, by image analysis of IgCa, IgG2b, IgG3, IgA, IgM, and IgE antibodies; and (2) to investigate the functional correlates of Ig structure by image analysis of antibody-complement complexes and Ig variants. The proposed research will be carried out on a collaborative basis by members of three laboratories, and will employ hybridoma, two-dimensional (2D) crystallization, and monolayer X-ray diffraction techniques. Isotype switch variants will be produced from existing hybridomas. Novel antibody variants will be generated by genetic engineering techniques and transformation with expression vectors. The variants will include Ig's with modified, deleted, and rearranged domains. Variants will be crystallized in 2D monolayers of lipid haptens and imaged at 15 resolution by electron microscopy and computer processing. The results should be informative about the shape, dimensions, domain structure, and hinge angles of the antibody molecules. Crystals of IgG's will be complexed with components of complement and imaged to reveal the structural basis of antibody-complement interaction. The monolayer X-ray diffraction technique will be developed and applied to obtain information missing from the electron microscope image analysis.
| Oberthuer, Dominik; Knoška, Juraj; Wiedorn, Max O et al. (2017) Double-flow focused liquid injector for efficient serial femtosecond crystallography. Sci Rep 7:44628 |
| Eagen, Kyle P; Aiden, Erez Lieberman; Kornberg, Roger D (2017) Polycomb-mediated chromatin loops revealed by a subkilobase-resolution chromatin interaction map. Proc Natl Acad Sci U S A 114:8764-8769 |
| Robinson, Philip J; Trnka, Michael J; Bushnell, David A et al. (2016) Structure of a Complete Mediator-RNA Polymerase II Pre-Initiation Complex. Cell 166:1411-1422.e16 |
| Lorch, Yahli; Kornberg, Roger D (2015) Chromatin-remodeling and the initiation of transcription. Q Rev Biophys 48:465-70 |
| Guan, Shenheng; Trnka, Michael J; Bushnell, David A et al. (2015) Deconvolution method for specific and nonspecific binding of ligand to multiprotein complex by native mass spectrometry. Anal Chem 87:8541-6 |
| Eagen, Kyle P; Hartl, Tom A; Kornberg, Roger D (2015) Stable Chromosome Condensation Revealed by Chromosome Conformation Capture. Cell 163:934-46 |
| Robinson, Philip J; Trnka, Michael J; Pellarin, Riccardo et al. (2015) Molecular architecture of the yeast Mediator complex. Elife 4: |
| Fazal, Furqan M; Meng, Cong A; Murakami, Kenji et al. (2015) Real-time observation of the initiation of RNA polymerase II transcription. Nature 525:274-7 |
| Lu, Jonathan; Trnka, Michael J; Roh, Soung-Hun et al. (2015) Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes. J Am Soc Mass Spectrom 26:2141-51 |
| Murakami, Kenji; Mattei, Pierre-Jean; Davis, Ralph E et al. (2015) Uncoupling Promoter Opening from Start-Site Scanning. Mol Cell 59:133-8 |
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