Abstract

This proposal represents a unified and internally self-contained effort to solve the structure of a few specific cell membrane proteins by high resolution electron crystallography. Research during the 5-year period of this grant will focus on bacteriorhodopsin and the various intermediates in its photocycle, PhoE porin, LamB porin, the adenine nucleotide exchanged protein from mitochondria, and a plasma membrane proton ATPase. The Program possesses all technical expertise and research facilities required for isolation and purification of the desired proteins, for biophysical and biochemical characterization (and modification, as desired), for reconstitution as two-dimensional crystalline sheets and for crystallographic structure analysis. Collaboration with expert investigators at other institutions is however, pursued when duplication of effort would be wasteful and unproductive.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
2P01GM036884-04
Application #
3096249
Study Section
(SSS)
Project Start
1986-07-01
Project End
1994-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
4
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Lawrence Berkeley National Laboratory
Department
Type
Organized Research Units
DUNS #
078576738
City
Berkeley
State
CA
Country
United States
Zip Code
94720

  Publications

Burkard, F; Chen, F; Kuziemko, G M et al. (1997) Electron density projection map of the botulinum neurotoxin 900-kilodalton complex by electron crystallography. J Struct Biol 120:78-84
Wolf, S G; Nogales, E; Kikkawa, M et al. (1996) Interpreting a medium-resolution model of tubulin: comparison of zinc-sheet and microtubule structure. J Mol Biol 262:485-501
Hud, N V; Downing, K H; Balhorn, R (1995) A constant radius of curvature model for the organization of DNA in toroidal condensates. Proc Natl Acad Sci U S A 92:3581-5
Perkins, G A; Downing, K H; Glaeser, R M (1995) Crystallographic extraction and averaging of data from small image areas. Ultramicroscopy 60:283-94
Nogales, E; Wolf, S G; Zhang, S X et al. (1995) Preservation of 2-D crystals of tubulin for electron crystallography. J Struct Biol 115:199-208
Han, B G; Wolf, S G; Vonck, J et al. (1994) Specimen flatness of glucose-embedded biological materials for electron crystallography is affected significantly by the choice of carbon evaporation stock. Ultramicroscopy 55:1-5
Han, B G; Vonck, J; Glaeser, R M (1994) The bacteriorhodopsin photocycle: direct structural study of two substrates of the M-intermediate. Biophys J 67:1179-86
Vonck, J; Han, B G; Burkard, F et al. (1994) Two progressive substrates of the M-intermediate can be identified in glucose-embedded, wild-type bacteriorhodopsin. Biophys J 67:1173-8
Wolf, S G; Mosser, G; Downing, K H (1993) Tubulin conformation in zinc-induced sheets and macrotubes. J Struct Biol 111:190-9
Perkins, G A; Burkard, F; Liu, E et al. (1993) Glucose alone does not completely hydrate bacteriorhodopsin in glucose-embedded purple membrane. J Microsc 169:61-5

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