Bacteriorhodopsin mutants of Halobacterium sp. GRB. I. The 5-bromo-2'-deoxyuridine selection as a method to isolate point mutants in halobacteria
- Max-Planck-Institut fuer Biochemie, Martinsried (West Germany)
Halobacterium sp. GRB was used to isolate bacteriorhodopsin (BR) mutants. A procedure is described which allows the enrichment of any type of mutant unable to grow under the selection conditions applied. Its use for the isolation of phototrophically negative, retinal-positive mutants of Halobacterium sp. GRB is demonstrated. Single-cell clones of this phenotype were further characterized. The expression of bacterioopsin was tested with a monoclonal antibody directed against the C terminus of the protein. The expressed bacteriorhodopsins were characterized by their specific activity for proton pumping, their spectral properties, and photocycle kinetics. About 15 independent mutants carrying bacteriorhodopsins of three distinct phenotypic classes could be isolated, including BR with a different absorption maximum, BR of lower specific activity, and BR characterized by a slower photocycle and a lack of proton pumping activity.
- OSTI ID:
- 5607805
- Journal Information:
- Journal of Biological Chemistry; (USA), Journal Name: Journal of Biological Chemistry; (USA) Vol. 264:22; ISSN JBCHA; ISSN 0021-9258
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ANTIBODIES
ANTIMETABOLITES
AZINES
BROMOURACILS
BUDR
CHEMICAL COMPOSITION
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
MONOCLONAL ANTIBODIES
MUTATIONS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHENOTYPE
PHOTOSYNTHETIC BACTERIA
PIGMENTS
PROTEINS
PYRIMIDINES
RHODOPSIN
RIBOSIDES
URACILS