Abstract

Starting from the firm foundation of the high resolution structure of the MscL channel provided by Rees and coworkers, this proposal weeks to develop new ion channels with novel functions and properties. In addition, this component of the PPG will develop several general capabilities in support of the other projects. The explicit goals are: 1. Establish capabilities for thorough characterization of MscL 2. Develop a new, higher-throughput assay for MscL channel gating 3. Computer modeling of MscL 4. Structure/function studies of MscL 5. Identify open channel blockers 6. Develop MscL derivatives with new gating properties This work will serve a dual purpose. First, we will develop the necessary skills to efficiently characterize MscL and related structures, in anticipation of the large number of new structures that will be prepared in this proposal and in Projects 1 and 3. In addition, we will use MscL as a platform to probe in detail the relationships between molecular structure and channel function. We will also modify the basic MscL channel to incorporate new properties such as ion selectively or gating in response to external stimuli.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM062532-03
Application #
6651783
Study Section
Special Emphasis Panel (ZRG1)
Project Start
2002-09-01
Project End
2003-08-31
Budget Start
Budget End
Support Year
3
Fiscal Year
2002
Total Cost
Indirect Cost

  Institution

Name
California Institute of Technology
Department
Type
DUNS #
078731668
City
Pasadena
State
CA
Country
United States
Zip Code
91125

  Publications

Elmore, Donald E (2010) Bringing ion channel crystal structures into sharper focus with computer modeling: examples from mechanosensitive channels. Future Med Chem 2:909-13
Maurer, Joshua A; Elmore, Donald E; Clayton, Daniel et al. (2008) Confirming the revised C-terminal domain of the MscL crystal structure. Biophys J 94:4662-7
Maurer, Joshua A; White, Victor E; Dougherty, Dennis A et al. (2007) Reconstitution of ion channels in agarose-supported silicon orifices. Biosens Bioelectron 22:2577-84
Spronk, Steven A; Elmore, Donald E; Dougherty, Dennis A (2006) Voltage-dependent hydration and conduction properties of the hydrophobic pore of the mechanosensitive channel of small conductance. Biophys J 90:3555-69
Kochendoerfer, Gerd G; Clayton, Daniel; Becker, Christian (2005) Chemical synthesis approaches to the engineering of ion channels. Protein Pept Lett 12:737-41
Kochendoerfer, Gerd G; Jones, David H; Lee, Sangwon et al. (2004) Functional characterization and NMR spectroscopy on full-length Vpu from HIV-1 prepared by total chemical synthesis. J Am Chem Soc 126:2439-46
Clayton, Daniel; Shapovalov, George; Maurer, Joshua A et al. (2004) Total chemical synthesis and electrophysiological characterization of mechanosensitive channels from Escherichia coli and Mycobacterium tuberculosis. Proc Natl Acad Sci U S A 101:4764-9
Becker, Christian F W; Strop, Pavel; Bass, Randal B et al. (2004) Conversion of a mechanosensitive channel protein from a membrane-embedded to a water-soluble form by covalent modification with amphiphiles. J Mol Biol 343:747-58
Becker, Christian F W; Clayton, Daniel; Shapovalov, George et al. (2004) On-resin assembly of a linkerless lanthanide(III)-based luminescence label and its application to the total synthesis of site-specifically labeled mechanosensitive channels. Bioconjug Chem 15:1118-24
Shapovalov, George; Lester, Henry A (2004) Gating transitions in bacterial ion channels measured at 3 microns resolution. J Gen Physiol 124:151-61

Showing the most recent 10 out of 19 publications