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Title: Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H + -ATPase

Authors:
ORCiD logo; ; ; ; ORCiD logo
Publication Date:
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1400000
Grant/Contract Number:
DEFG02-88ER13938; 1410164; 1713899; P30EY016665
Resource Type:
Journal Article: Published Article
Journal Name:
Plant Physiology (Bethesda)
Additional Journal Information:
Journal Name: Plant Physiology (Bethesda); Journal Volume: 176; Journal Issue: 1; Related Information: CHORUS Timestamp: 2018-01-12 13:58:32; Journal ID: ISSN 0032-0889
Publisher:
American Society of Plant Biologists (ASPB)
Country of Publication:
United States
Language:
English

Citation Formats

Haruta, Miyoshi, Tan, Li Xuan, Bushey, Daniel B., Swanson, Sarah J., and Sussman, Michael R. Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H + -ATPase. United States: N. p., 2017. Web. doi:10.1104/pp.17.01126.
Haruta, Miyoshi, Tan, Li Xuan, Bushey, Daniel B., Swanson, Sarah J., & Sussman, Michael R. Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H + -ATPase. United States. doi:10.1104/pp.17.01126.
Haruta, Miyoshi, Tan, Li Xuan, Bushey, Daniel B., Swanson, Sarah J., and Sussman, Michael R. Tue . "Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H + -ATPase". United States. doi:10.1104/pp.17.01126.
@article{osti_1400000,
title = {Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H + -ATPase},
author = {Haruta, Miyoshi and Tan, Li Xuan and Bushey, Daniel B. and Swanson, Sarah J. and Sussman, Michael R.},
abstractNote = {},
doi = {10.1104/pp.17.01126},
journal = {Plant Physiology (Bethesda)},
number = 1,
volume = 176,
place = {United States},
year = {Tue Oct 17 00:00:00 EDT 2017},
month = {Tue Oct 17 00:00:00 EDT 2017}
}

Journal Article:
Free Publicly Available Full Text
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  • H/sup +/-ATPase activity in plasma membranes isolated from Avena sativa root cells is inhibited by N-ethylmaleimide, a covalent modifier of protein sulfhydryl groups. The rate of inhibition is reduced by ADP, MgADP, and MgATP, but even at 40 millimolar ADP the enzyme is only partially protected against inactivation. When plasma membranes are treated with N-(2-/sup 3/H)ethylmaleimide and analyzed by sodium dodecyl sulfate polyaerylamide gel electrophoresis, prominent radioactive bands appear at M/sub r/ = 100,000 and several other positions. However, only radioactivity in the M/sub r/ = 100,000 protein is reduced by the presence of MgADP. These results provide independent evidencemore » that the M/sub r/ = 100,000 polypeptide which is observed in purified preparations of the enzyme is the catalytic subunit of the H/sup +/-ATPase. When tryptic peptides are produced from N-(2-/sup 3/H)ethylmaleimide labeled M/sub r/ = 100,000 protein and separated by reverse phase high performance liquid chromatography, two radioactive peaks are observed for which N-(2-/sup 3/H)ethylmaleimide incorporation is reduced in the presence of MgADP.« less
  • The molecular weight and isoelectric point of the plasma membrane H/sup +/-ATPase from red beet storage tissue were determined using N,N'-dicyclohexylcarbodiimide (DCCD) and a H/sup +/-ATPase antibody. When plasma membrane vesicles were incubated with 20 micromolar (/sup 14/C)-DCCD at 0/sup 0/C, a single 97,000 dalton protein was visualized on a fluorography of a sodium dodecyl sulfate polyacrylamide gel. A close correlation between (/sup 14/C)DCCD labeling of the 97,000 dalton protein and the extent of ATPase inhibition over a range of DCCD concentration suggests that this 97,000 dalton protein is a component of the plasma membrane H/sup +/-ATPase. An antibody raisedmore » against the plasma membrane H/sup +/-ATPase of Neurospora crassa cross-reacted with the 97,000 dalton DCCD-binding protein, further supporting the identity of this protein. Immunoblots of two-dimensional gels of red beet plasma membrane vesicles indicated the isoelectric point of the H/sup +/-ATPase to be 6.5.« less
  • The carboxyl-modifying reagent N,N'-dicyclohexylcarbodiimide (DCCD) inactivates the ATPase with pseudo-first order kinetics, suggesting that one site on the enzyme is involved. The rate constant for inactivation at pH 7.5 and 30/sup 0/C is approximately 1000 M/sup -1/ min/sup -1/, similar to values reported for the DCCD-binding proteolipid of F/sub 0/-F/sub 1/-type (H/sup +/)-ATPases and for the sarcoplasmic reticulum (Ca/sup +2/)-ATPase. Although hydrophobic carbodiimides are inhibitory at micromolar concentrations, a hydrophilic analogue, 1-ethyl-3-(dimethylaminopropyl)-carbodiimide, is completely inactive even at millimolar concentrations. This result implies that the DCCD-reactive site is located in a lipophilic environment. (/sup 14/C)DCCD is incorporated into the M/sub r/more » = 104,000 polypeptide at a rate similar to the rate of inactivation. There is no evidence for a separate low molecular weight DCCD-binding proteolipid. Using quantitative amino acid analysis, we established that complete inhibition occurs at a stoichiometry of 0.4 mol of DCCD/mol of polypeptide. Overall, the results are consistent with the idea the DCCD reacts with a single amino acid residue of the Neuspora (H/sup +/)-ATPase, thereby blcoking ATP hydrolysis and proton translocation. 21 references, 5 figures, 2 tables.« less
  • The carboxyl group activating reagent N-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline (EEDQ) interacts with the Neurospora plasma membrane H/sup +/-ATPase in at least three different ways. This reagent irreversibly inhibits ATP hydrolysis with kinetics that are pseudo-first-order at several concentrations of EEDQ, and an appropriate transform of these data suggests that 1 mol of EEDQ inactivates 1 mol of the H/sup +/-ATPase. Inhibition probably involves activation of an ATPase carboxyl group followed by a nucleophilic attack by a vicinal nucleophilic functional group in the ATPase polypeptide chain, resulting in an intra-molecular cross-link. The enzyme is protected against EEDQ inhibition by MgATP in the presence ofmore » vanadate, a combination of ligands that has previously been shown to lock the H/sup +/-APTase in a conformation that presumably resembles the transition states of the enzyme phosphorylation and dephosphorylation reactions, but is not protected by the substrate analogue MgADP. The ATPase is also labeled by the exogenous nucleophile (/sup 14/C)glycine ethyl ester in an EEDQ-dependent reaction, and the labeling is diminished in the presence of MgATP plus vanadate. In a third type of interaction, EEDQ mediates the specific cross-linking of ATPase monomers with some other membrane protein, possibly another ATPase monomer, leading to the formation of a product with an apparent molecular weight of about 260,000.« less
  • In plants, the transport of solutes across the plasma membrane is driven by a proton pump (H{sup +}-ATPase) that produces an electric potential and pH gradient. The authors isolated and sequenced a full-length cDNA clone that encodes this enzyme in Arabidopsis thaliana. The protein predicted from its nucleotide sequence encodes 959 amino acids and has a molecular mass of 104,207 Da. The plant protein shows structural features common to a family of cation-translocating ATPases found in the plasma membrane of prokaryotic and eukaryotic cells, with the greatest overall identity in amino acid sequence (36%) to the H{sup +}-ATPase observed inmore » the plasma membrane of fungi. The structure predicted from a hydropathy plant contains at least eight transmembrane segments, with most of the protein (73%) extending into the cytoplasm and only 5% of the residues exposed on the external surface. Unique features of the plant enzyme include diverged sequences at the amino and carboxyl termini as well as greater hydrophilic character in three extracellular loops.« less