Over the last year, we have made progress in several areas that are at the focus of work in our laboratory. Specific progress includes: (i) determination of the spatial and molecular architecture of the E.coli apparatus for chemotaxis using advanced methods in cryo electron tomography,(ii) development and application of methods for direct structure determination of membrane proteins in intact cells by combining electron crystallography and cryo electron tomography of cells under near-native conditions,(iii) determination of the structure of intact HIV and SIV virions using novel approaches that combine high-throughput electron tomography with advanced tools for image analysis,(iv) discovery and structural analysis of the HIV entry claw that is formed prior to viral entry into T-cells,(v) establishment of powerful methods for automated image segmentation that can cope with the very low signal-to-noise ratios in the electron tomograms recorded in both room temperature and cryo tomographic experiments, and (vi) development of a novel technology for automated cell and tissue imaging at 100 nm resolution using dual beam electron microscopy.

National Institute of Health (NIH)
Division of Basic Sciences - NCI (NCI)
Intramural Research (Z01)
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Basic Sciences
United States
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Fenton, Andrew K; Hobley, Laura; Butan, Carmen et al. (2010) A coiled-coil-repeat protein 'Ccrp' in Bdellovibrio bacteriovorus prevents cellular indentation, but is not essential for vibroid cell morphology. FEMS Microbiol Lett 313:89-95
Milne, Jacqueline L S; Subramaniam, Sriram (2009) Cryo-electron tomography of bacteria: progress, challenges and future prospects. Nat Rev Microbiol 7:666-75
Lengyel, Jeffrey S; Milne, Jacqueline L S; Subramaniam, Sriram (2008) Electron tomography in nanoparticle imaging and analysis. Nanomedicine (Lond) 3:125-31
Bartesaghi, A; Sprechmann, P; Liu, J et al. (2008) Classification and 3D averaging with missing wedge correction in biological electron tomography. J Struct Biol 162:436-50
Narasimha, Rajesh; Aganj, Iman; Bennett, Adam E et al. (2008) Evaluation of denoising algorithms for biological electron tomography. J Struct Biol 164:7-17
Lefman, Jonathan; Morrison, Robert; Subramaniam, Sriram (2007) Automated 100-position specimen loader and image acquisition system for transmission electron microscopy. J Struct Biol 158:318-26
Subramaniam, Sriram; Bartesaghi, Alberto; Liu, Jun et al. (2007) Electron tomography of viruses. Curr Opin Struct Biol 17:596-602
Heymann, Jurgen A W; Hayles, Mike; Gestmann, Ingo et al. (2006) Site-specific 3D imaging of cells and tissues with a dual beam microscope. J Struct Biol 155:63-73
Subramaniam, Sriram (2006) The SIV surface spike imaged by electron tomography: one leg or three? PLoS Pathog 2:e91
Milne, Jacqueline L S; Wu, Xiongwu; Borgnia, Mario J et al. (2006) Molecular structure of a 9-MDa icosahedral pyruvate dehydrogenase subcomplex containing the E2 and E3 enzymes using cryoelectron microscopy. J Biol Chem 281:4364-70

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