Characterization of the cytochrome c oxidase in isolated and purified plasma membranes from the cyanobacterium Anacystis nidulans
- Univ. of Vienna (Austria)
Functionally intact plasma membranes were isolated from the cyanobacterium (blue-green alga) Anacystis nidulans through French pressure cell extrusion of lysozyme/EDTA-treated cells, separated from thylakoid membranes by discontinuous sucrose density gradient centrifugation, and purified by repeated recentrifugation. Origin and identity of the chlorophyll-free plasma membrane fraction were confirmed by labeling of intact cells with impermeant protein markers, ({sup 35}S)diazobenzenesulfonate and fluorescamine, prior to membrane isolation. Rates of oxidation of reduced horse heart cytochrome c by purified plasma and thylakoid membranes were 90 and 2 nmol min{sup {minus}1} (mg of protein){sup {minus}1}, respectively. The cytochrome oxidase in isolated plasma membranes was identified as a copper-containing aa{sub 3}-type enzyme from the properties of its redox-active and EDTA-resistant Cu{sup 2+} ESR signal, the characteristic inhibition profile, reduced minus oxidized difference spectra, carbon monoxide difference spectra, photoaction and photodissociation spectra of the CO-inhibited enzyme, and immunological cross-reaction of two subunits of the enzyme with antibodies against subunits I and II, and the holoenzyme, of Paracoccus denitrificans aa{sub 3}-type cytochrome oxidase. The data presented are the first comprehensive evidence for the occurrence of aa{sub 3}-type cytochrome oxidase in the plasma membrane of a cyanobacterium similar to the corresponding mitochondrial enzyme.
- DOE Contract Number:
- FG03-87ER13736
- OSTI ID:
- 5478585
- Journal Information:
- Biochemistry; (USA), Vol. 28:7; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CYANOBACTERIA
ENZYME ACTIVITY
CYTOCHROME OXIDASE
MOLECULAR STRUCTURE
CARBON MONOXIDE
CELL MEMBRANES
CYTOCHROMES
ELECTRON SPIN RESONANCE
ELECTROPHORESIS
OXIDATION
PURIFICATION
SULFUR 35
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON COMPOUNDS
CARBON OXIDES
CELL CONSTITUENTS
CHALCOGENIDES
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
ENZYMES
EVEN-ODD NUCLEI
HAEM DEHYDROGENASES
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MEMBRANES
MICROORGANISMS
NUCLEI
ORGANIC COMPOUNDS
OXIDES
OXIDOREDUCTASES
OXYGEN COMPOUNDS
PIGMENTS
PROTEINS
RADIOISOTOPES
RESONANCE
SULFUR ISOTOPES
550201* - Biochemistry- Tracer Techniques