The NCMI is dedicated to the advancement of cryo-electron microscopy and tomography methodology for structure determination of macromolecules, molecular machines and cells in their various functional states at the highest possible resolutions. Having completed 10 C-alpha backbone traceable cryo-EM structures of molecular machines during the current grant period, we are well poised to tackle the next set of challenging structural biology problems. Our technology research development will focus around optimization of cutting edge instrumentation, data collection strategy, data management, image processing, modeling and visualization from electron images recorded from two unique combinations of instrumentation: (i) a 300 kV electron microscope with a direct electron detector and an in-column energy filter and (ii) a 200 kV electron microscope with a direct electron detector, an in-column energy filter and a Zernike phase contrast optics. Our technology development is led by 10 driving biological projects and also synergizes with 10 user projects that together include animal, plant and bacterial viruses, apoptosis-causing protein machine, chaperonin-substrate complex, membrane ion channels, nuclear receptor-coactivator complex, small RNA, oncogene protein complex, lipoproteins, amyloid protein aggregates, neuronal cells, virus-infected cells and mammalian cells related to cancer and eye diseases. Specifically, our Center will focus on 3 technology research and development projects: (i) Characterize and determine the optimal utilization of direct electron detectors and Zernike phase optics; (ii) Extend the structural determinations of biochemically purified molecular machines beyond current resolution limits; (iii) Develop novel methodologies for analyzing subcellular structures i cells by cryo-ET. We will proactively identify new projects from NIH-funded investigators across the US. We will continue our rigor in disseminating our software, experimental and computational protocols via workshops and web seminars. We will maintain an engaging advisory board to critique our progress and guide our strategic planning annually. Our efforts will extend cryo-EM/cryo-ET capabilities to fill the information gaps between x-ray crystallography, NMR and optical microscopy from nanometer to atomic resolutions.

Public Health Relevance

Our proposed cryo-electron microscopy and tomography methodology is targeted to study structures of biologically active macromolecules, molecular machines and cells, which are potential drug targets for treating or preventing diseases. Our projects cover specimens relevant to infectious diseases (viruses and bacteria), neurodegenerative diseases (chaperonins and amyloid), eye disease (rod cell), cancer (complexes involved in gene expression and signaling); cardiovascular diseases (lipoprotein and ion channels).

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Biotechnology Resource Grants (P41)
Project #
2P41GM103832-30
Application #
8743504
Study Section
Special Emphasis Panel (ZRG1-OBT-A (40))
Program Officer
Swain, Amy L
Project Start
1996-12-01
Project End
2019-12-13
Budget Start
2015-02-17
Budget End
2015-12-31
Support Year
30
Fiscal Year
2015
Total Cost
$1,884,602
Indirect Cost
$658,909
Name
Baylor College of Medicine
Department
Biochemistry
Type
Schools of Medicine
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
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Pintilie, Grigore; Chiu, Wah (2018) Assessment of structural features in Cryo-EM density maps using SSE and side chain Z-scores. J Struct Biol 204:564-571
Qi, Xiaodong; Zhang, Fei; Su, Zhaoming et al. (2018) Programming molecular topologies from single-stranded nucleic acids. Nat Commun 9:4579
Dai, Wei; Chen, Muyuan; Myers, Christopher et al. (2018) Visualizing Individual RuBisCO and Its Assembly into Carboxysomes in Marine Cyanobacteria by Cryo-Electron Tomography. J Mol Biol 430:4156-4167

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