Steady-state ATP synthesis by bacteriorhodopsin and chloroplast coupling factor co-reconstituted into asolectin vesicles.

Journal Article (Journal Article)

A method was developed for the co-reconstitution of bacteriorhodopsin and chloroplast coupling factor in asolectin vesicles. First, bacteriorhodopsin was reconstituted from a mixture of octyl glucoside, asolectin, and protein in the presence of ethylenediaminetetraacetic acid by passage through a Sephadex G-50 centrifuge column. Then, the purified coupling factor was reconstituted from a mixture of sodium cholate, bacteriorhodopsin vesicles, and coupling factor in the presence of Mg2+ by passage through the centrifuge column. Sucrose density-gradient centrifugation indicated a band of vesicles with slightly different positions in the gradient for maximum vesicle concentration, bacteriorhodopsin vesicle concentration, ATP synthesis, and ATP hydrolysis. The rate of light-driven ATP synthesis reaches a limiting value as the concentration of bacteriorhodopsin and the light intensity are increased. A steady-state rate of ATP synthesis of 1 mumol per mg of coupling factor X min-1 has been achieved. Apparently this rate is limited by the heterogeneity within the vesicle population and by the ability of bacteriorhodopsin to form a sufficiently large pH gradient.

Full Text

Duke Authors

Cited Authors

  • Krupinski, J; Hammes, GG

Published Date

  • June 1986

Published In

Volume / Issue

  • 83 / 12

Start / End Page

  • 4233 - 4237

PubMed ID

  • 2872676

Pubmed Central ID

  • PMC323706

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.83.12.4233


  • eng

Conference Location

  • United States