Partial purification of the chloroplast ATP synthase from Chlamydomonas reinhardtii and the cloning and sequencing of a cDNA encoding the gamma subunit
The chloroplast ATP synthase was partially purified from the green alga Chlamydomonas reinhardtii by extracting membranes with deoxycholate and KCl, followed by centrifugation and ammonium sulfate fractionation of the supernatant. The enzyme assay involved the reconstitution of such fractions with bacteriorhodopsin and soybean phospholipids to form vesicles capable of light-dependent ({sup 32}P)-phosphate esterification. A cDNA for the gamma subunit from Chlamydomonas was isolated, expressed in vitro and sequenced. It contains the entire coding region for the gamma subunit precursor. A 35 amino acid long transit peptide resides at the NH{sub 2}-terminus of a 323 amino acid long mature peptide that is 77% similar to the spinach gamma subunit. Six cysteines were found; three were conserved in Chlamydomonas and spinach.
- Research Organization:
- Wisconsin Univ., Madison, WI (USA)
- OSTI ID:
- 7245998
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
HYDROLASES
FRACTIONATION
AMINO ACID SEQUENCE
CELL MEMBRANES
CENTRIFUGATION
CHLAMYDOMONAS
CHLOROPLASTS
CYSTEINE
DNA SEQUENCING
DNA-CLONING
ESTERIFICATION
PHOSPHOLIPIDS
PHOSPHORUS 32
RHODOPSIN
SPINACH
TRACER TECHNIQUES
ALGAE
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHLOROPHYCOTA
CLONING
DAYS LIVING RADIOISOTOPES
DNA HYBRIDIZATION
ENZYMES
ESTERS
FOOD
HYBRIDIZATION
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIPIDS
MAGNOLIOPHYTA
MAGNOLIOPSIDA
MEMBRANES
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHORUS ISOTOPES
PIGMENTS
PLANTS
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
STRUCTURAL CHEMICAL ANALYSIS
THIOLS
UNICELLULAR ALGAE
VEGETABLES
550201* - Biochemistry- Tracer Techniques