scholarly journals Inhibition of the H+ -ATPase in bovine adrenal chromaffin granule ghosts by diethylstilbestrol Evidence for a tight coupling between ATP hydrolysis and proton translocation

FEBS Letters ◽  
1988 ◽  
Vol 229 (1) ◽  
pp. 40-44 ◽  
Author(s):  
Martin Grønberg ◽  
Torgeir Flatmark
Keyword(s):  
Atp Hydrolysis ◽  
Proton Translocation ◽  
Chromaffin Granule ◽  
Tight Coupling ◽  
Chromaffin Granule Ghosts ◽  
Adrenal Chromaffin

scholarly journals Isolation of ATPase I, the proton pump of chromaffin-granule membranes

1985 ◽  
Vol 231 (3) ◽  
pp. 557-564 ◽  
Author(s):  
J M Percy ◽  
J G Pryde ◽  
D K Apps
Keyword(s):  
Proton Pump ◽  
Alkylating Agents ◽  
Proton Translocation ◽  
Phase Segregation ◽  
Chromaffin Granule ◽  
Chromaffin Granule Ghosts ◽  
Triton X ◽  
Atpase Ii ◽  
Hydrolysis Of ◽  
Granule Matrix

Chromaffin-granule membranes contain two ATPases, which can be separated by (NH4)2SO4 fractionation after solubilization with detergents, or by phase segregation in Triton X-114. ATPase I (Mr 400000) is inhibited by trialkyltin, quercetin and alkylating agents, and hydrolyses both ATP and ITP. It contains up to five types of subunit, including a low-Mr hydrophobic polypeptide that reacts with dicyclohexylcarbodi-imide; these subunits are unrelated to those of mitochondrial F1F0-ATPase, as judged by size and reaction with antibodies. ATPase II (Mr 140000) is inhibited by vanadate, and is specific for ATP; it has not been extensively purified. Proton translocation by resealed chromaffin-granule ‘ghosts’, measured by uptake of methylamine or by quenching of the fluorescence of 9-amino-6-chloro-2-methoxyacridine, is supported by the hydrolysis of ATP or ITP, and inhibited by quercetin or alkylating agents, but not by vanadate. ATPase I must therefore be the proton translocator involved in the uptake of catecholamines and possibly of other components of the chromaffin-granule matrix, whereas ATPase II does not translocate protons.

scholarly journals Inhibition of adenosine triphosphatase, 5-hydroxytryptamine transport and proton-translocation activities of resealed chromaffin-granule ‘ghosts’

1980 ◽  
Vol 190 (2) ◽  
pp. 273-282 ◽  
Author(s):  
David K. Apps ◽  
James G. Pryde ◽  
Raul Sutton ◽  
John H. Phillips
Keyword(s):  
Molecular Weight ◽  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Proton Translocation ◽  
Low Molecular Weight ◽  
Chromaffin Granule ◽  
Low Concentrations ◽  
Tributyltin Chloride ◽  
Granule Membrane ◽  
Chromaffin Granule Ghosts

1. Highly purified resealed chromaffin-granule ‘ghosts’ were assayed for ATPase and ATP-driven H+-translocation and 5-hydroxytryptamine-uptake activities, and for 5-hydroxytryptamine uptake driven by an imposed transmembrane H+-gradient. The effects of several inhibitors on these activities were studied. 2. Dicyclohexylcarbodi-imide inhibits all of these activities, but not in parallel; at low concentrations it decreases the permeability of the membrane to protons. 3. 4-Chloro-7-nitrobenzofuran (Nbf-Cl) and silicotungstate inhibit ATP-dependent activities, without effect on 5-hydroxytryptamine uptake driven by an imposed H+-gradient. 4. Tributyltin chloride inhibits all of the activities. 5. Treatment of the ‘ghosts’ with low concentrations of urea inhibits 5-hydroxytryptamine uptake and ATP-dependent generation of a transmembrane H+-gradient, without inhibiting ATPase activity. 6. Nbf-Cl and silicotungstate are without effect on the rate of leakage of 5-hydroxytryptamine from preloaded ‘ghosts’, whereas dicyclohexylcarbodi-imide and tributyltin chloride accelerate the rate of leakage. 7. Treatment of the membranes with 14C-labelled Nbf-Cl labels several proteins; membranes treated with dicyclohexyl[14C]carbodi-imide are labelled predominantly in a protein of low molecular weight, which may be analogous to the mitochondrial H+-conducting proteolipid. 8. It is concluded that Nbf-Cl and silicotungstate inhibit the H+-translocating ATPase of the granule membrane; that dicyclohexylcarbodi-imide inhibits the ATPase, and inhibits 5-hydroxytryptamine accumulation by accelerating leakage of the amine; and that the effects of tributyltin chloride are due to inhibition of the ATPase, and collapse of the transmembrane H+-gradient through OH−-anion exchange.

scholarly journals Proton translocation of the bovine chromaffin-granule membrane

1978 ◽  
Vol 170 (3) ◽  
pp. 661-672 ◽  
Author(s):  
J H Phillips ◽  
V P Allison
Keyword(s):  
Atp Hydrolysis ◽  
Proton Translocation ◽  
Potential Gradient ◽  
Protonmotive Force ◽  
Chromaffin Granule ◽  
Anion Transporter ◽  
Direct Measurements ◽  
Granule Membrane ◽  
Potential Gradients ◽  
Proton Movement

Bovine chromaffin granules were lysed and their membranes resealed to give osmotically sensitive ‘ghosts’. These swell in the presence of salts and MgATP. It is shown that this is due to proton entry accompanied by anions. The rate of swelling depends on the anion present, but swelling is not limited to media containing permeant anions. It is quite marked in solutions of sulphates, phosphates and acetates. It is not uncoupler-sensitive, suggesting that at least one component of swelling is due to coupled proton and anion entry (non-electrogenic proton translocation). Direct measurements of transmembrane pH and potential gradients generated in the presence of MgATP shows that these are rapidly established in sucrose media, and are rather little affected by the presence of salts. They contribute roughly equally to the total protonmotive force. The potential gradient is establihsed very rapidly, but the pH gradient is generated over several minutes. The gradients are not completely dissipated by uncoupler, and it is shown that, in media containing sulphate but no permeant anion, sulphate can be taken up by the ‘ghosts’. There thus appear to be two mechanisms of proton translocation across the membrane, both dependent on ATP hydrolysis: an electrogenic transfer of protons, and proton movement linked to an anion transporter of broad specificity.

scholarly journals ATP-dependent proton translocation in resealed chromaffin granule ghosts

FEBS Letters ◽  
1977 ◽  
Vol 78 (1) ◽  
pp. 53-56 ◽  
Author(s):  
Torgeir Flatmark ◽  
Ole Chr. Ingebretsen
Keyword(s):  
Proton Translocation ◽  
Chromaffin Granule ◽  
Chromaffin Granule Ghosts

scholarly journals Vacuolar H+-ATPase of adrenal secretory granules. Rapid partial purification and reconstitution into proteoliposomes

1990 ◽  
Vol 271 (1) ◽  
pp. 127-131 ◽  
Author(s):  
J R Perez-Castiñeira ◽  
D K Apps
Keyword(s):  
Atp Hydrolysis ◽  
Secretory Granules ◽  
Gel Filtration ◽  
Partial Purification ◽  
Weak Base ◽  
Chromaffin Granule ◽  
Octyl Glucoside ◽  
Rapid Purification ◽  
Adrenal Chromaffin ◽  
Triton X

A procedure has been developed for the rapid purification and reconstitution into phospholipid vesicles of the proton-translocating ATPase of bovine adrenal chromaffin-granule membranes. It involves fractionation of the membranes with Triton X-114, resolubilization of the ATPase with n-octyl glucoside, addition of purified lipids and removal of detergent by gel filtration. The entire process can be completed within 2 h. H+ translocation was detected by the ATP-dependent quenching of the fluorescence of a permeant weak base. The effect of varying the lipid composition of the vesicles on ATP hydrolysis and H+ translocation by the reconstituted enzyme was examined. ATPase activity was maximally increased about 4-fold by added lipid, but was relatively insensitive to its composition, whereas vesicle acidification was absolutely dependent on the addition of phospholipids and cholesterol.

CRISPR/Cas9-Mediated Gene Editing of Vacuolar ATPase Subunit D Mediates Phytohormone Biosynthesis and Virus Resistance in Rice

2021 ◽  
Author(s):  
Qinghua Lu ◽  
Xiangwen Luo ◽  
Xiao Yang ◽  
Tong Zhou ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Virus Resistance ◽  
Atp Hydrolysis ◽  
Proton Translocation ◽  
Resistance Breeding ◽  
Rice Stripe Virus ◽  
Proton Pumps ◽  
Wild Type ◽  
Knockout Mutant ◽  
Atpase Subunit ◽  
Vacuolar Atpases

Abstract Background: Vacuolar ATPases (v-ATPases) are proton pumps for proton translocation across membranes that utilize energy derived from ATP hydrolysis; Previous research revealed Osv-ATPases mediates phytohormes levels and resistance in rice. Osv-ATPase subunit d (Osv-ATPase d) is part of an integral, membrane-embedded V0 complex of V-ATPases complex, whether Osv-ATPase d involves in phytohormes biosynthesis and resistance in rice remains unknown.Finding: The knockout mutant line (line 5) of Osv-ATPase d was generated using the CRISPR/Cas9 system, mutation of Osv-ATPase d did not show any detrimental effect on plant growth or yield productivity. Transcriptomic results showed Osv-ATPase d probably involved in mediating the biosynthesis of plant hormones and resistance in rice. Mutation of Osv-ATPase d significantly increased JA and ABA biosynthesis than wild type. Compared to wild type, mutation of Osv-ATPase d increased the resistance against Southern rice black-streaked dwarf virus (SRBSDV), however, decreased the resistance against Rice stripe virus (RSV) in rice. Conclusion: Taken together, our data reveal the Osv-ATPase d mediates phytohormone biosynthesis and virus resistance in rice, which can be selected as a potential target for resistance breeding in rice.

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