Our objectives are to determine the three-dimensional (3D) crystal structures of macromolecules at near atomic resolution and the 3-D structures of non-crystalline macromolecular assemblies at intermediate resolution, by a combination of electron microscopy and computer processing methods. The present proposal is a resource related project. Requests include the acquisition of a medium high voltage (300 kV) electron microscope with appropriate accessories and an operational fund to develop the technology for the above aims. Technical development will be focused on specimen preparation, 3-D data collection at low dose and low temperature conditions and implementation of computer programs for 3-D reconstruction. The developed technology will be immediately applied to the high resolution crystal structure determination of crotoxin complex, T4 DNA helix destabilizing protein, fungus lysozyme and Fc fragments; and to the intermediate resolution structure determination of muscle Z disc and flagella axoneme. Collaborative projects will be established with investigators outside our research groups after the midpoint of the grant period. We plan to complete a thorough analysis of the potential as well as limitations of this technology in its general applicability to a variety of biological systems. We expect to present a final recommendation on the feasibility and desirability of establishing a national shared facility of this technology at the end of this grant period.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR002250-02
Application #
3104086
Study Section
(SSS)
Project Start
1985-03-01
Project End
1989-02-28
Budget Start
1986-03-01
Budget End
1987-02-28
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Arizona
Department
Type
Schools of Arts and Sciences
DUNS #
City
Tucson
State
AZ
Country
United States
Zip Code
85722
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