Factors defining the functional coupling of bacteriorhodopsin and ATP synthase in liposomes

R.L. Van Der Bend*, J.B.W.J. Cornelissen, J.A. Berden, K. Van Dam

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

25 Citations (Scopus)

Abstract

Optimal conditions for the co-reconstitution of bacteriorhodopsin and yeast mitochondria ATP synthase were determined. Reconstitution was achieved with a quick two-step procedure. Preparations obtained by this method displayed in optimal cases 2-3-times higher activities (up to 500 nmol ATP/min per mg protein) compared with maximal values reported in the literature, when light-driven ATP synthesis was measured under similar conditions. The final activities depended on the purification method used for the ATP synthase, and it is shown that the oligomycin-sensitive ATP hydrolysis activity was not a good measure for the ability of the ATP synthase preparations to perform ATP synthesis after co-reconstitution. Light-driven ATP synthesis activities depended also on the type of phospholipid used, soybean phospholipid giving the best results. A close relation to the bacteriorhodopsin proton pump activity was found. Using different phospholipids, different H+ ATP ratios were found, calculated from ATP synthesis activities and initial and steady-state light-driven proton pump activities. From this, together with the findings that the ATP synthase displayed the same ATP hydrolysis and ATP-32Pi exchange activities with these different phospholipids used, it is concluded that the protein distribution for the two proteins among the liposomes is different relative to each other for the different phospholipids. The light-driven ATP synthesis activity did not correlate with the variation in leakiness of the membrane for protons when different phospholipids were used. An explanation is given by the finding that at high light intensities, the ATP synthesis became independent of the presence of protonophore.

Original languageEnglish
Pages (from-to)87-101
JournalBBA - Bioenergetics
Volume767
Issue number1
DOIs
Publication statusPublished - 26 Oct 1984
Externally publishedYes

Keywords

  • ATP synthase
  • Bacteriorhodopsin
  • Co-reconstitution
  • Liposomes

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