A Conformational Change of the γ Subunit Indirectly Regulates the Activity of Cyanobacterial F1-ATPase

Sunamura, Ei-Ichiro; Konno, Haruyoshi; Imashimizu, Mari; Mochimaru, Mari; Hisabori, Toru

doi:10.1074/jbc.m112.395053

Cited by 11 publications

(16 citation statements)

References 54 publications

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“…9), an interaction with βI397 was not required for changed ADP inhibition and redox regulation upon mutation. Some contradictions in our study might be attributed to an additional parameter that was described recently and could play a role in the CF 1 -ATPase redox regulation in general [32]. Therein, the authors demonstrated that restricting relative slippage of the γ-termini diminishes ADP inhibition.…”

Section: F 1-redox Mutant Enzyme Effect On Atpase Redox Regulationcontrasting

confidence: 57%

“…The methods applied here were slightly modified from previous reports [30,32]. The complex was purified from E. coli cell extracts in Buffer A, which contained 20 mM HEPES-KOH (pH 8.0), 100 mM KCl, 0.1 mM MgCl 2 , 0.1 mM ADP, and 50 mM imidazole.…”

Section: Expression and Purification Of The α 3 β 3 γ Complexmentioning

confidence: 99%

“…Activity measurements were carried out as previously described [32], except for phosphoenolpyruvate, which was used at 0.2 mM.…”

Section: Measurement Of Atp Hydrolysis Activitymentioning

confidence: 99%

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Redox regulation of CF1-ATPase involves interplay between the γ-subunit neck region and the turn region of the βDELSEED-loop

Buchert

Konno

Hisabori

2015

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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The soluble F1 complex of ATP synthase (FoF1) is capable of ATP hydrolysis, accomplished by the minimum catalytic core subunits α3β3γ. A special feature of cyanobacterial F1 and chloroplast F1 (CF1) is an amino acid sequence inserted in the γ-subunit. The insertion is extended slightly into the CF1 enzyme containing two additional cysteines for regulation of ATPase activity via thiol modulation. This molecular switch was transferred to a chimeric F1 by inserting the cysteine-containing fragment from spinach CF1 into a cyanobacterial γ-subunit [Y. Kim et al., redox regulation of rotation of the cyanobacterial F1-ATPase containing thiol regulation switch, J Biol Chem, 286 (2011) 9071-9078]. Under oxidizing conditions, the obtained F1 tends to lapse into an ADP-inhibited state, a common regulation mechanism to prevent wasteful ATP hydrolysis under unfavorable circumstances. However, the information flow between thiol modulation sites on the γ-subunit and catalytic sites on the β-subunits remains unclear. Here, we clarified a possible interplay for the CF1-ATPase redox regulation between structural elements of the βDELSEED-loop and the γ-subunit neck region, i.e., the most convex part of the α-helical γ-termini. Critical residues were assigned on the β-subunit, which received the conformation change signal produced by disulfide/dithiol formation on the γ-subunit. Mutant response to the ATPase redox regulation ranged from lost to hypersensitive. Furthermore, mutant cross-link experiments and inversion of redox regulation indicated that the γ-redox state might modulate the subunit interface via reorientation of the βDELSEED motif region.

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Section: F 1-redox Mutant Enzyme Effect On Atpase Redox Regulationcontrasting

confidence: 57%

Section: Expression and Purification Of The α 3 β 3 γ Complexmentioning

confidence: 99%

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Redox regulation of CF1-ATPase involves interplay between the γ-subunit neck region and the turn region of the βDELSEED-loop

Buchert

Konno

Hisabori

2015

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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“…The structure of both the plastid and mitochondrial ATP synthases has been studied extensively (Hamasur and Glaser, 1992;Yoshida et al, 2001;Sunamura et al, 2012). We have shown previously that the subunits from both complexes turn over at comparable rates in Arabidopsis cell culture .…”

Section: Atp Synthasesmentioning

confidence: 99%

Proteins with High Turnover Rate in Barley Leaves Estimated by Proteome Analysis Combined with in Planta Isotope Labeling

et al. 2014

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Protein turnover is a key component in cellular homeostasis; however, there is little quantitative information on degradation kinetics for individual plant proteins. We have used 15 N labeling of barley (Hordeum vulgare) plants and gas chromatography-mass spectrometry analysis of free amino acids and liquid chromatography-mass spectrometry analysis of proteins to track the enrichment of 15 N into the amino acid pools in barley leaves and then into tryptic peptides derived from newly synthesized proteins. Using information on the rate of growth of barley leaves combined with the rate of degradation of 14 N-labeled proteins, we calculate the turnover rates of 508 different proteins in barley and show that they vary by more than 100-fold. There was approximately a 9-h lag from label application until 15 N incorporation could be reliably quantified in extracted peptides. Using this information and assuming constant translation rates for proteins during the time course, we were able to quantify degradation rates for several proteins that exhibit half-lives on the order of hours. Our workflow, involving a stringent series of mass spectrometry filtering steps, demonstrates that 15 N labeling can be used for large-scale liquid chromatography-mass spectrometry studies of protein turnover in plants. We identify a series of abundant proteins in photosynthesis, photorespiration, and specific subunits of chlorophyll biosynthesis that turn over significantly more rapidly than the average protein involved in these processes. We also highlight a series of proteins that turn over as rapidly as the well-known D1 subunit of photosystem II. While these proteins need further verification for rapid degradation in vivo, they cluster in chlorophyll and thiamine biosynthesis.

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“…These values are comparable with those of other inhibited bacterial F-ATPases where no inhibitor protein is involved. For example, the values for the F 1 -ATPase from the cyanobacterium Thermosynechococcus elongatus are 0.2 and 9.0 U per milligram of protein before and after activation with LDAO (Sunamura et al, 2012). For C. thermarum they are 0.9 and 28.5 U per milligram of protein before and after activation (Keis et al, 2006), and for the chloroplast F 1 -ATPase from Spinacia oleracea they are 4.4 and 39.7 U per milligram of protein before and after activation (Groth & Schirwitz, 1999).…”

Section: Resultsmentioning

confidence: 99%

Structure of a catalytic dimer of the α- and β-subunits of the F-ATPase fromParacoccus denitrificansat 2.3 Å resolution

Morales-Ríos

Montgomery

Leslie

et al. 2015

Acta Crystallogr F Struct Biol Commun

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The structures of F-ATPases have predominantly been determined from mitochondrial enzymes, and those of the enzymes in eubacteria have been less studied.Paracoccus denitrificansis a member of the α-proteobacteria and is related to the extinct protomitochondrion that became engulfed by the ancestor of eukaryotic cells. TheP. denitrificansF-ATPase is an example of a eubacterial F-ATPase that can carry out ATP synthesis only, whereas many others can catalyse both the synthesis and the hydrolysis of ATP. Inhibition of the ATP hydrolytic activity of theP. denitrificansF-ATPase involves the ζ inhibitor protein, an α-helical protein that binds to the catalytic F1domain of the enzyme. This domain is a complex of three α-subunits and three β-subunits, and one copy of each of the γ-, δ- and ∊-subunits. Attempts to crystallize the F1–ζ inhibitor complex yielded crystals of a subcomplex of the catalytic domain containing the α- and β-subunits only. Its structure was determined to 2.3 Å resolution and consists of a heterodimer of one α-subunit and one β-subunit. It has no bound nucleotides, and it corresponds to the `open' or `empty' catalytic interface found in other F-ATPases. The main significance of this structure is that it aids in the determination of the structure of the intact membrane-bound F-ATPase, which has been crystallized.

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A Conformational Change of the γ Subunit Indirectly Regulates the Activity of Cyanobacterial F1-ATPase

Cited by 11 publications

References 54 publications

Redox regulation of CF1-ATPase involves interplay between the γ-subunit neck region and the turn region of the βDELSEED-loop

Redox regulation of CF1-ATPase involves interplay between the γ-subunit neck region and the turn region of the βDELSEED-loop

Proteins with High Turnover Rate in Barley Leaves Estimated by Proteome Analysis Combined with in Planta Isotope Labeling

Structure of a catalytic dimer of the α- and β-subunits of the F-ATPase fromParacoccus denitrificansat 2.3 Å resolution

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