ATP synthesis catalyzed by the ATPase complex from Rhodospirillum rubrum reconstituted into phospholipid vesicles together with bacteriorhodopsin

Oren, Rachel; Weiss, Sara; Garty, Haim; Caplan, S. Roy; Gromet-Elhanan, Zippora

doi:10.1016/0003-9861(80)90133-2

Cited by 21 publications

(4 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is possible that ATP ^P¡ exchange includes combined reactions, catalyzed by adenylate kinase (ATP 5=± ADP exchange) and polynucleotide phosphorylase (ADP -P¡ exchange). Very similar exchange reactions were observed in the partially purified preparations of F0F[ from R. rubrum (Oren et al, 1980). ATP ^P¡ exchange was nearly completely inhibited by AP5A in the purified preparations of 0.…”

Section: Methodssupporting

confidence: 74%

Empirical estimation of interaction energies for ligands binding in the isolated .beta.-subunit of F0F1 ATP synthase from Rhodospirillum rubrum

Khananshvili¹

1988

Biochemistry

View full text Add to dashboard Cite

show abstract

Section: Methodssupporting

confidence: 74%

Empirical estimation of interaction energies for ligands binding in the isolated .beta.-subunit of F0F1 ATP synthase from Rhodospirillum rubrum

Khananshvili¹

1988

Biochemistry

View full text Add to dashboard Cite

show abstract

“…The maximum level of light-dependent ATP synthesis in those studies was -100 nmol-mg-1 min-1. This value is comparable to that observed in a variety of co-reconstituted preparations (8)(9)(10)(11)(12). Recently, van der Bend et al (13) reported synthetic rates of up to 500 nmol-mg-l min-' with co-reconstituted preparations of the mitochondrial ATP synthase.…”

supporting

confidence: 62%

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

Krupinski¹,

Hammes²

1986

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

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 ofsodium 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 Ismol per mg of coupling factor-min 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.Bacteriorhodopsin (bR) is a light-driven proton pump found in the purple membrane of Halobacterium halobium (cf. ref. 1). It is readily reconstituted into phospholipid vesicles in which the interior is acidified upon illumination (cf. ref. 2). Racker and Stoeckenius (3) were the first to demonstrate that vesicles co-reconstituted with bR and the ATP synthesizing complex, initially from mitochondria, synthesized ATP from ADP and inorganic phosphate upon illumination of the vesicles. The observed rates of ATP synthesis were -0.1% of the rates for oxidative phosphorylation found in mitochondria. A highly active co-reconstituted preparation of bR and ATP synthase would be an ideal model system with which to perform studies on the mechanism of energy coupling. Consequently, this experimental system has been refined and applied to other ATPases.The dicyclohexylcarbodiimide-sensitive ATP-synthesizing complex from chloroplasts (DSA) (cf. ref. 4) was first successfully co-reconstituted with bR by Winget et al. (5). Subsequently, the co-reconstituted system was used to study the steady-state kinetics of ATP synthesis and hydrolysis (6,7). The maximum level of light-dependent ATP synthesis in those studies was -100 nmol-mg-1 min-1. This value is comparable to that observed in a variety of co-reconstituted preparations (8-12). Recently, van der Bend et al. (13) reported synthetic rates of up to 500 nmol-mg-l min-' with co-reconstituted preparations of the mitochondrial ATP synthase. In the present study, we report the co-reconstitution of bR and DSA from spinach chloroplasts with maximum rates of light-driven ATP synthesis of -1 ,umol-mg-1-min-. MATERIALS AND METHODSChemicals. Asolectin was from Associated Concentrates (Woodside, NY). Sephadex G-50 fine was from Pharmacia Fine Chemicals. Pyranine was from Molecular Probes (Plano, TX). Octyl B-D-glucopyranoside was from Ca...

show abstract

“…The ATP synthase from Rsp. rubrum has been isolated (Oren & Gromet-Elhanan, 1977; Bengis-Garber & Gromet-Elhanan, 1979) and characterized biochemically (Oren et al, 1980). It is similar to other ATP synthases, being a multi-subunit enzyme consisting of two structurally and functionally distinct sectors called Fl and Fo.…”

Section: Introductionmentioning

confidence: 99%

DNA sequence of a gene cluster coding for subunits of the F0 membrane sector of ATP synthase in Rhodospirillum rubrum. Support for modular evolution of the F1 and F0 sectors

Falk

Walker

1988

Biochemical Journal

View full text Add to dashboard Cite

A region was cloned from the genome of the purple non-sulphur photobacterium Rhodospirillum rubrum that contains genes coding for the membrane protein subunits of the F0 sector of ATP synthase. The clone was identified by hybridization with a synthetic oligonucleotide designed on the basis of the known protein sequence of the dicyclohexylcarbodi-imide-reactive proteolipid, or subunit c. The complete nucleotide sequence of 4240 bp of this region was determined. It is separate from an operon described previously that encodes the five subunits of the extrinsic membrane sector of the enzyme, F1-ATPase. It contains a cluster of structural genes encoding homologues of all three membrane subunits a, b and c of the Escherichia coli ATP synthase. The order of the genes in Rsp. rubrum is a-c-b'-b where b and b' are homologues. A similar gene arrangement for F0 subunits has been found in two cyanobacteria, Synechococcus 6301 and Synechococcus 6716. This suggests that the ATP synthase complexes of all these photosynthetic bacteria contain nine different polypeptides rather than eight found in the E. coli enzyme; the chloroplast ATP synthase complex is probably similar to the photosynthetic bacterial enzymes in this respect. The Rsp. rubrum b subunit is modified after translation. As shown by N-terminal sequencing of the protein, the first seven amino acid residues are removed before or during assembly of the ATP synthase complex. The subunit-a gene is preceded by a gene coding for a small hydrophobic protein, as has been observed previously in the atp operons in E. coli, bacterium PS3 and cyanobacteria. A number of features suggest that the Rsp. rubrum cluster of F0 genes is an operon. On its 5' side are found sequences resembling the -10 (Pribnow) and -35 boxes of E. coli promoters, and the gene cluster is followed by a sequence potentially able to form a stable stem-loop structure, suggesting that it acts as a rho-independent transcription terminator. These features and the small intergenic non-coding sequences suggest that the genes are cotranscribed, and so the name atp2 is proposed for this second operon coding for ATP synthase subunits in Rsp. rubrum. The finding that genes for the F0 and F1 sectors of the enzyme are in separate clusters supports the view that these represent evolutionary modules.

show abstract

ATP synthesis catalyzed by the ATPase complex from Rhodospirillum rubrum reconstituted into phospholipid vesicles together with bacteriorhodopsin

Cited by 21 publications

References 28 publications

Empirical estimation of interaction energies for ligands binding in the isolated .beta.-subunit of F0F1 ATP synthase from Rhodospirillum rubrum

Empirical estimation of interaction energies for ligands binding in the isolated .beta.-subunit of F0F1 ATP synthase from Rhodospirillum rubrum

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

DNA sequence of a gene cluster coding for subunits of the F0 membrane sector of ATP synthase in Rhodospirillum rubrum. Support for modular evolution of the F1 and F0 sectors

Contact Info

Product

Resources

About