Reconstitution of halorhodopsin
Halobacterium halobium contains a family of retinal-bound proteins: bacteriorhodopsin (bR) which mediates phototrophic growth as a light-riven proton pump, halorhodopsin (hR) which is a light-driven chloride pump, and one or more sensory rhodopsins (sR) which mediate a phototactic response. Two-dimensional crystallization of halorhodopsin has been attempted though the reconstitution of purified halorhodopsin with purple membrane lipid for electron microscopy work. The first important step for crystallization is to get a homogeneous protein which is pure and not denatured. Homogeneous halorhodopsin has been obtained by a modification of existing purification methods. Some nice looking membrane patches which have the same density as purple membrane have been obtained. But unfortunately, they are not crystalline. The procedure of hR reconstitution is described in detail and some other strategies to induce the protein crystal in the reconstituted membrane are discussed in this dissertation. 76 refs., 20 figs., 6 tabs.
- Research Organization:
- Lawrence Berkeley Lab., CA (USA)
- Sponsoring Organization:
- DOE/ER; DOHHS
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 6374468
- Report Number(s):
- LBL-28624; ON: DE91001791; CNN: GM36884
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ATP
BACTERIA
BRINES
CHEMISTRY
CRYSTALLIZATION
DICHROISM
ELECTRON MICROSCOPY
MICROORGANISMS
MICROSCOPY
NUCLEOTIDES
ORGANIC COMPOUNDS
PHASE TRANSFORMATIONS
PHOTOCHEMISTRY
PIGMENTS
PROTEINS
PURIFICATION
RHODOPSIN
SPECTROSCOPY