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      The plasma membrane H +‐ATPase, a simple polypeptide with a long history

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          Abstract

          The plasma membrane H +‐ATPase of fungi and plants is a single polypeptide of fewer than 1,000 residues that extrudes protons from the cell against a large electric and concentration gradient. The minimalist structure of this nanomachine is in stark contrast to that of the large multi‐subunit F OF 1 ATPase of mitochondria, which is also a proton pump, but under physiological conditions runs in the reverse direction to act as an ATP synthase. The plasma membrane H +‐ATPase is a P‐type ATPase, defined by having an obligatory phosphorylated reaction cycle intermediate, like cation pumps of animal membranes, and thus, this pump has a completely different mechanism to that of F OF 1 ATPases, which operates by rotary catalysis. The work that led to these insights in plasma membrane H +‐ATPases of fungi and plants has a long history, which is briefly summarized in this review.

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          Crystal structure of the plasma membrane proton pump.

          A prerequisite for life is the ability to maintain electrochemical imbalances across biomembranes. In all eukaryotes the plasma membrane potential and secondary transport systems are energized by the activity of P-type ATPase membrane proteins: H+-ATPase (the proton pump) in plants and fungi, and Na+,K+-ATPase (the sodium-potassium pump) in animals. The name P-type derives from the fact that these proteins exploit a phosphorylated reaction cycle intermediate of ATP hydrolysis. The plasma membrane proton pumps belong to the type III P-type ATPase subfamily, whereas Na+,K+-ATPase and Ca2+-ATPase are type II. Electron microscopy has revealed the overall shape of proton pumps, however, an atomic structure has been lacking. Here we present the first structure of a P-type proton pump determined by X-ray crystallography. Ten transmembrane helices and three cytoplasmic domains define the functional unit of ATP-coupled proton transport across the plasma membrane, and the structure is locked in a functional state not previously observed in P-type ATPases. The transmembrane domain reveals a large cavity, which is likely to be filled with water, located near the middle of the membrane plane where it is lined by conserved hydrophilic and charged residues. Proton transport against a high membrane potential is readily explained by this structural arrangement.
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            Energization of transport processes in plants. roles of the plasma membrane H+-ATPase.

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              Molecular characterization of mutant Arabidopsis plants with reduced plasma membrane proton pump activity.

              Arabidopsis mutants containing gene disruptions in AHA1 and AHA2, the two most highly expressed isoforms of the Arabidopsis plasma membrane H(+)-ATPase family, have been isolated and characterized. Plants containing homozygous loss-of-function mutations in either gene grew normally under laboratory conditions. Transcriptome and mass spectrometric measurements demonstrate that lack of lethality in the single gene mutations is not associated with compensation by increases in RNA or protein levels. Selected reaction monitoring using synthetic heavy isotope-labeled C-terminal tryptic peptides as spiked standards with a triple quadrupole mass spectrometer revealed increased levels of phosphorylation of a regulatory threonine residue in both isoforms in the mutants. Using an extracellular pH assay as a measure of in vivo ATPase activity in roots, less proton secreting activity was found in the aha2 mutant. Among 100 different growth conditions, those that decrease the membrane potential (high external potassium) or pH gradient (high external pH) caused a reduction in growth of the aha2 mutant compared with wild type. Despite the normal appearance of single mutants under ideal laboratory growth conditions, embryos containing homozygous double mutations are lethal, demonstrating that, as expected, this protein is absolutely essential for plant cell function. In conclusion, our results demonstrate that the two genes together perform an essential function and that the effects of their single mutations are mostly masked by overlapping patterns of expression and redundant function as well as by compensation at the post-translational level.
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                Author and article information

                Contributors
                palmgren@plen.ku.dk
                Journal
                Yeast
                Yeast
                10.1002/(ISSN)1097-0061
                YEA
                Yeast (Chichester, England)
                John Wiley and Sons Inc. (Hoboken )
                0749-503X
                1097-0061
                10 December 2018
                April 2019
                : 36
                : 4 , André Goffeau ( doiID: 10.1002/yea.v36.4 )
                : 201-210
                Affiliations
                [ 1 ] Department of Plant and Environmental Sciences University of Copenhagen Frederiksberg C Denmark
                [ 2 ] Louvain Institute of Biomolecular Science and Technology (LIBST) UCLouvain Louvain‐la‐Neuve Belgium
                Author notes
                [*] [* ] Correspondence

                Michael Palmgren, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, DK‐1871 Frederiksberg C, Denmark.

                Email: palmgren@ 123456plen.ku.dk

                Author information
                https://orcid.org/0000-0002-9982-6114
                https://orcid.org/0000-0001-7780-7230
                Article
                YEA3365 YEA-Oct-18-0104.R1
                10.1002/yea.3365
                6590192
                30447028
                3f63c880-2afb-412d-81a1-4573de559100
                © 2018 The Authors. Yeast published by John Wiley & Sons, Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 16 October 2018
                : 08 November 2018
                : 11 November 2018
                Page count
                Figures: 0, Tables: 0, Pages: 10, Words: 6272
                Categories
                Special Issue Article
                Special Issue Articles
                Custom metadata
                2.0
                yea3365
                April 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.4 mode:remove_FC converted:24.06.2019

                Molecular biology
                arabidopsis thaliana,f‐type atpase,neurospora crassa,nicotiana tabacum,proton pump,p‐type atpase,saccharomyces cerevisiae,schizosaccharomyces pombe

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