Activity, amount and subunit composition of vacuolar-type H+-ATPase and H+-PPase in wheat roots under severe NaCl stress
Identifieur interne : 002F72 ( Main/Exploration ); précédent : 002F71; suivant : 002F73Activity, amount and subunit composition of vacuolar-type H+-ATPase and H+-PPase in wheat roots under severe NaCl stress
Auteurs : B. S. Wang [République populaire de Chine] ; Rafael Ratajczak [Allemagne] ; J. H. Zhang [Hong Kong]Source :
- Journal of Plant Physiology [ 0176-1617 ] ; 2000.
English descriptors
- KwdEn :
- ATP, CAM, DTT, EGTA, Hepes, P-ATPase, PBS, PMSF, PVP, Tricine, Triticum aestivum, V-ATPase, V-PPase, immunoprecipitation, severe NaCl stress, wheat root.
Abstract
Summary: Tonoplast-enriched membrane vesicles from wheat (Triticum aestivum) roots exposed to severe NaCl stress (200 mmol/L) for 3 days were isolated using the Dextran T70 step gradient method. High purity of the tonoplast vesicle preparations was indicated by a high degree of Bafilomycin A1 and nitrate inhibition of ATP-hydrolysis activity at pH 7.5. Severe NaCl stress strongly reduced the V-ATPase and V-PPase substrate hydrolysis activity compared with controls. Immunoprecipitation analysis showed that the relative amount of V-ATPase protein related to total tonoplast proteins was reduced by a factor of 2.4 due to NaCl stress. Antisera directed against the Kalanchoë daigremontiana V-ATPase holoenzyme cross-reacted with polypeptides present in the T. aestivum root tonoplast vesicle fraction exhibiting apparent molecular masses of 67, 58, 44, 41, 34, 32 and 17kDa, which are likely to be subunits of the wheat V-ATPase. In preparations from salt-treated seedlings an additional 33 kDa polypeptide was immunodecorated by an antiserum against V-ATPase subunit A, which is discussed to represent a proteolytic fragment of subunit A. By cross-reaction with a specific antiserum a 70 kDa polypeptide was identified as V-PPase. Similar to V-ATPase subunits the V-PPase protein amount was lower in preparations from NaCl-treated plants.
Url:
DOI: 10.1016/S0176-1617(00)80143-1
Affiliations:
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Wang</name><affiliation wicri:level="1"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Biology Department, Shandong Teachers' University</wicri:regionArea><wicri:noRegion>Shandong Teachers' University</wicri:noRegion></affiliation></author><author><name sortKey="Ratajczak, Rafael" sort="Ratajczak, Rafael" uniqKey="Ratajczak R" first="Rafael" last="Ratajczak">Rafael Ratajczak</name><affiliation wicri:level="3"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Botany, Darmstadt University of Technology, Fachbereich 10, Schnittspahnstraße 3-5, D-64287 Darmstadt</wicri:regionArea><placeName><region type="land" nuts="1">Hesse (Land)</region><region type="district" nuts="2">District de Darmstadt</region><settlement type="city">Darmstadt</settlement></placeName></affiliation><affiliation></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Allemagne</country></affiliation></author><author><name sortKey="Zhang, J H" sort="Zhang, J H" uniqKey="Zhang J" first="J. 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Zhang</name><affiliation wicri:level="1"><country xml:lang="fr">Hong Kong</country><wicri:regionArea>Department of Biology, Hong Kong Baptist University, Kowloon Tong</wicri:regionArea><wicri:noRegion>Kowloon Tong</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Plant Physiology</title><title level="j" type="abbrev">JPLPH</title><idno type="ISSN">0176-1617</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2000">2000</date><biblScope unit="volume">157</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="109">109</biblScope><biblScope unit="page" to="116">116</biblScope></imprint><idno type="ISSN">0176-1617</idno></series><idno type="istex">CEDA5715608AE36978393017F72DECFDD31B28FB</idno><idno type="DOI">10.1016/S0176-1617(00)80143-1</idno><idno type="PII">S0176-1617(00)80143-1</idno><idno type="ArticleID">80143</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0176-1617</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>ATP</term><term>CAM</term><term>DTT</term><term>EGTA</term><term>Hepes</term><term>P-ATPase</term><term>PBS</term><term>PMSF</term><term>PVP</term><term>Tricine</term><term>Triticum aestivum</term><term>V-ATPase</term><term>V-PPase</term><term>immunoprecipitation</term><term>severe NaCl stress</term><term>wheat root</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Summary: Tonoplast-enriched membrane vesicles from wheat (Triticum aestivum) roots exposed to severe NaCl stress (200 mmol/L) for 3 days were isolated using the Dextran T70 step gradient method. High purity of the tonoplast vesicle preparations was indicated by a high degree of Bafilomycin A1 and nitrate inhibition of ATP-hydrolysis activity at pH 7.5. Severe NaCl stress strongly reduced the V-ATPase and V-PPase substrate hydrolysis activity compared with controls. Immunoprecipitation analysis showed that the relative amount of V-ATPase protein related to total tonoplast proteins was reduced by a factor of 2.4 due to NaCl stress. Antisera directed against the Kalanchoë daigremontiana V-ATPase holoenzyme cross-reacted with polypeptides present in the T. aestivum root tonoplast vesicle fraction exhibiting apparent molecular masses of 67, 58, 44, 41, 34, 32 and 17kDa, which are likely to be subunits of the wheat V-ATPase. In preparations from salt-treated seedlings an additional 33 kDa polypeptide was immunodecorated by an antiserum against V-ATPase subunit A, which is discussed to represent a proteolytic fragment of subunit A. By cross-reaction with a specific antiserum a 70 kDa polypeptide was identified as V-PPase. Similar to V-ATPase subunits the V-PPase protein amount was lower in preparations from NaCl-treated plants.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>Hong Kong</li><li>République populaire de Chine</li></country><region><li>District de Darmstadt</li><li>Hesse (Land)</li></region><settlement><li>Darmstadt</li></settlement></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Wang, B S" sort="Wang, B S" uniqKey="Wang B" first="B. S." last="Wang">B. S. Wang</name></noRegion></country><country name="Allemagne"><region name="Hesse (Land)"><name sortKey="Ratajczak, Rafael" sort="Ratajczak, Rafael" uniqKey="Ratajczak R" first="Rafael" last="Ratajczak">Rafael Ratajczak</name></region><name sortKey="Ratajczak, Rafael" sort="Ratajczak, Rafael" uniqKey="Ratajczak R" first="Rafael" last="Ratajczak">Rafael Ratajczak</name></country><country name="Hong Kong"><noRegion><name sortKey="Zhang, J H" sort="Zhang, J H" uniqKey="Zhang J" first="J. H." last="Zhang">J. H. Zhang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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