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Calcium transport in vesicles energized by cytochrome oxidase

Thesis/Dissertation ·
DOI:https://doi.org/10.2172/6252979· OSTI ID:6252979
 [1]
  1. Univ. of Rochester, NY (United States)

Experiments on the reconstitution of cytochrome oxidase into phospholipid vesicles were carried out using techniques of selectivity energizing the suspensions with ascorbate and cytochrome c or ascorbate, PMS, and internally trapped cytochrome c. It was found that the K+ selective ionophore valinomycin stimulated the rate of respiration of cytochrome oxidase vesicles regardless of the direction of the K+ flux across the vesicle membranes. The stimulation occurred in the presence of protonophoric uncouplers and in the complete absence of potassium or in detergent-lysed suspensions. Gramicidin had similar effects and it was determined that the ionophores acted by specific interaction with cytochrome oxidase rather than by the previously assumed collapse of membrane potentials. When hydrophobic proteins and appropriate coupling factors were incorporated into the cytochrome oxidase, vesicles phosphorylation of ADP could be coupled to the oxidation reaction of cytochrome oxidase. Relatively low P:O, representing poor coupling of the system, were problematical and precluded measurements of protonmotive force. However the system was used to study ion translocation.

Research Organization:
Univ. of Rochester, NY (United States). Dept. of Radiation Biology and Biophysics
Sponsoring Organization:
USDOE
DOE Contract Number:
EY-76-C-02-3490
OSTI ID:
6252979
Report Number(s):
UR--3490-1549(Vols.1-2)
Country of Publication:
United States
Language:
English