Use of inside-out chloroplast thylakoid membrane vesicles for studying electron transport and membrane structure

Thesis/Dissertation · Thu Jan 01 00:00:00 EST 1987

OSTI ID:5718764

Atta-Asafo-Adjei, E

Inside-out and right-side-out thylakoid vesicles were isolated from spinach chloroplasts by aqueous-polymer two-phase partitioning following mechanical fragmentation of thylakoid membranes by Yeda press treatment. Externally added plastocyanin stimulated the whole-chain and PSI electron transport rates in the inside-out thylakoid vesicles by about 500 and 350%, respectively, compared to about 50% stimulation for both assays in the fraction enriched in right-side-out vesicles. The electron transport between PSII and PSI in inside-out thylakoid vesicles appears to be interrupted due to plastocyanin release from the thylakoids by the Yeda press treatment, but it was restored by externally added plastocyanin. Acetic anhydride chemical modification and uncoupler-induced proton release from dark-adapted membranes are probes for detecting the sequested proton domains in thylakoid membranes. Both assays were used to find out if inside-out membranes retain metastable, localized proton binding domains. Treatment of dark-maintained inside-out thylakoid membrane vesicles with ({sup 3}H)acetic anhydride showed no uncoupler-induced increase in acetylation of the 33, 24, and 18 kDa polypeptides of the oxygen-evolving-complex, indicating complete loss of the implicated proton domains in these polypeptides. The various steps in the inside-out preparation were studied to discern which steps(s) leads to the loss of the metastable domain proton pool.

• Research Organization: Purdue Univ., Lafayette, IN (USA)
• OSTI ID: 5718764
• Resource Relation: Other Information: Thesis (Ph. D.)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PHOTOSYNTHETIC MEMBRANES
MOLECULAR STRUCTURE
PHOTOSYNTHETIC REACTION CENTERS
MEMBRANE TRANSPORT
ACETYLATION
CHLOROPLASTS
POLYPEPTIDES
PROTONS
QUERCETIN
SPINACH
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ACYLATION
AROMATICS
BARYONS
CELL CONSTITUENTS
CHEMICAL REACTIONS
ELEMENTARY PARTICLES
FERMIONS
FLAVENOIDS
FLAVONES
FOOD
HADRONS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
MAGNOLIOPHYTA
MAGNOLIOPSIDA
MEMBRANES
NUCLEONS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PEPTIDES
PHENOLS
PLANTS
POLYPHENOLS
PROTEINS
PYRANS
VEGETABLES
550201* - Biochemistry- Tracer Techniques
550301 - Cytology- Tracer Techniques