ATP synthase from a cyanobacterial <i>Synechocystis</i> 6803 mutant containing the regulatory segment of the chloroplast γ subunit shows thiol modulation

Krenn, Bea E.; Aardewijn, Pepijn; Walraven, H.S. van; Werner-Grüne, Sabine; Strotmann, Heinrich; Kraayenhof, Ruud

doi:10.1042/bst0230757

Cited by 16 publications

(10 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus the essential structural features and basic functions of both types of ␥ subunits should be the same. This is also consistent with previous reports that upon introduction of the chloroplast ␥-like regulatory stretch into the ␥ subunit of cyanobacterial F 1 conferred thiol modulation on this enzyme (23,24). In this mutant both ATP hydrolysis and ATP synthesis are controlled by thiol modulation.…”

Section: Discussionsupporting

confidence: 81%

“…In vitro reduction can be achieved by dithiothreitol (DTT) or other dithiols, but the natural reductant is reduced thioredoxin-f (Trx-f) (20 -22). Introduction of nine amino acids comprising the regulatory sequence into the cyanobacterial ␥ subunit induced thiol modulation in Synechocystis (23,24). Conversely, replacement of the two cysteines by serines in ␥ subunit of CF 0 CF 1 from Chlamydomonas reinhardtii (25,26) resulted in a non-modulated enzyme.…”

mentioning

confidence: 99%

See 1 more Smart Citation

ATPase Activity of a Highly Stable α3β3γ Subcomplex of Thermophilic F1 Can Be Regulated by the Introduced Regulatory Region of γ Subunit of Chloroplast F1

Bald

Noji²,

Stumpp

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

A mutant F 1 -ATPase ␣ 3 ␤ 3 ␥ subcomplex from the thermophilic Bacillus PS3 was constructed, in which 111 amino acid residues (Val 92 to Phe 202 ) from the central region of the ␥ subunit were replaced by the 148 amino acid residues of the homologous region from spinach chloroplast F 1 -ATPase ␥ subunit, including the regulatory stretch, and were designated as ␣ 3 ␤ 3 ␥ (TCT) (Thermophilic-Chloroplast-Thermophilic). By the insertion of this regulatory region into the ␥ subunit of thermophilic F 1 , we could confer the thiol modulation property to the thermophilic ␣ 3 ␤ 3 ␥ subcomplex. The overexpressed ␣ 3 ␤ 3 ␥ (TCT) was easily purified in large scale, and the ATP hydrolyzing activity of the obtained complex was shown to increase up to 3-fold upon treatment with chloroplast thioredoxin-f and dithiothreitol. No loss of thermostability compared with the wild type subcomplex was found, and activation by dithiothreitol was functional at temperatures up to 80°C. ␣ 3 ␤ 3 ␥ (TCT) was inhibited by the ⑀ subunit from chloroplast F 1 -ATPase but not by the one from the thermophilic F 1 -ATPase, indicating that the introduced amino acid residues from chloroplast F 1 -␥ subunit are important for functional interaction with the ⑀ subunit.

show abstract

Section: Discussionsupporting

confidence: 81%

mentioning

confidence: 99%

ATPase Activity of a Highly Stable α3β3γ Subcomplex of Thermophilic F1 Can Be Regulated by the Introduced Regulatory Region of γ Subunit of Chloroplast F1

Bald

Noji²,

Stumpp

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Interestingly, the normally redox-insensitive ATP synthase from Synechocystis sp. PCC 6803 was rendered sensitive to thiol modulation by mutagenesis inserting a cassette containing the plastid-type regulatory motif into its c-subunit (Krenn et al 1995;Werner-Grune et al 1994). Taken together, it may be concluded that the cyanobacterial Trxs must have functions quite different from those of the phylogenetically related chloroplast Trxs.…”

Section: Quantitative Analysis Of the Synechocystis Sp Pcc 6803 Trxsmentioning

confidence: 93%

The diversity and complexity of the cyanobacterial thioredoxin systems

et al. 2006

View full text Add to dashboard Cite

Cyanobacteria perform oxygenic photosynthesis, which makes them unique among the prokaryotes, and this feature together with their abundance and worldwide distribution renders them a central ecological role. Cyanobacteria and chloroplasts of plants and algae are believed to share a common ancestor and the modern chloroplast would thus be the remnant of an endosymbiosis between a eukaryotic cell and an ancestral oxygenic photosynthetic prokaryote. Chloroplast metabolic processes are coordinated with those of the other cellular compartments and are strictly controlled by means of regulatory systems that commonly involve redox reactions. Disulphide/dithiol exchange catalysed by thioredoxin is a fundamental example of such regulation and represents the molecular mechanism for light-dependent redox control of an ever-increasing number of chloroplast enzymatic activities. In contrast to chloroplast thioredoxins, the functions of the cyanobacterial thioredoxins have long remained elusive, despite their common origin. The sequenced genomes of several cyanobacterial species together with novel experimental approaches involving proteomics have provided new tools for re-examining the roles of the thioredoxin systems in these organisms. Thus, each cyanobacterial genome encodes between one and eight thioredoxins and all components necessary for the reduction of thioredoxins. Screening for thioredoxin target proteins in cyanobacteria indicates that assimilation and storage of nutrients, as well as some central metabolic pathways, are regulated by mechanisms involving disulphide/dithiol exchange, which could be catalysed by thioredoxins or related thiol-containing proteins.

show abstract

“…In addition, insertion of a ‘chloroplast‐like’ regulatory segment responsible for thiol modulation into the γ subunit of F 0 F 1 ‐ATPase of the cyanobacterium Synechocystis sp. PCC6803 at the chromosome level can be used to generate a redox‐sensitive F 0 F 1 , which is one of the main distinguishing features of the chloroplast‐type ATP synthase in vivo (Werner‐Grune et al , 1994; Krenn et al , 1995).…”

Section: Introductionmentioning

confidence: 99%

The regulator of the F1 motor: inhibition of rotation of cyanobacterial F1-ATPase by the ɛ subunit

Konno

Murakami-Fuse

Fujii

et al. 2006

EMBO J

View full text Add to dashboard Cite

The chloroplast-type F(1) ATPase is the key enzyme of energy conversion in chloroplasts, and is regulated by the endogenous inhibitor epsilon, tightly bound ADP, the membrane potential and the redox state of the gamma subunit. In order to understand the molecular mechanism of epsilon inhibition, we constructed an expression system for the alpha(3)beta(3)gamma subcomplex in thermophilic cyanobacteria allowing thorough investigation of epsilon inhibition. epsilon Inhibition was found to be ATP-independent, and different to that observed for bacterial F(1)-ATPase. The role of the additional region on the gamma subunit of chloroplast-type F(1)-ATPase in epsilon inhibition was also determined. By single molecule rotation analysis, we succeeded in assigning the pausing angular position of gamma in epsilon inhibition, which was found to be identical to that observed for ATP hydrolysis, product release and ADP inhibition, but distinctly different from the waiting position for ATP binding. These results suggest that the epsilon subunit of chloroplast-type ATP synthase plays an important regulator for the rotary motor enzyme, thus preventing wasteful ATP hydrolysis.

show abstract

ATP synthase from a cyanobacterial Synechocystis 6803 mutant containing the regulatory segment of the chloroplast γ subunit shows thiol modulation

Cited by 16 publications

References 10 publications

ATPase Activity of a Highly Stable α3β3γ Subcomplex of Thermophilic F1 Can Be Regulated by the Introduced Regulatory Region of γ Subunit of Chloroplast F1

ATPase Activity of a Highly Stable α3β3γ Subcomplex of Thermophilic F1 Can Be Regulated by the Introduced Regulatory Region of γ Subunit of Chloroplast F1

The diversity and complexity of the cyanobacterial thioredoxin systems

The regulator of the F1 motor: inhibition of rotation of cyanobacterial F1-ATPase by the ɛ subunit

Contact Info

Product

Resources

About