2005 Fiscal Year Final Research Report Summary
Molecular structure and functional mechanism of a novel proton pump H+-pyrophosphatase
| Project/Area Number |
13142203
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Nagoya University |
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Principal Investigator |
MAESHIMA Masayoshi Nagoya University, Graduate School of Bioagricultural Sciences, Professor, 大学院生命農学研究科, 教授 (80181577)
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| Co-Investigator(Kenkyū-buntansha) |
NAKANISHI Yoichi Nagoya University, Graduate School of Bioagricultural Sciences, Assistant Professor, 大学院生命農学研究科, 助手 (60362290)
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| Project Period (FY) |
2001 – 2005
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| Keywords | H+-pyrophosphatase / Proton pump / Higher-order structure / Vacuolar membrane / Membrane topology / Functional mechanism / エネルギー共役 / プロトンポンプ |
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| Research Abstract |
During the project period for five years, we obtained the following results concerning H+-pyrophosphatase and metal transporters.
(1) H^+-pyrophosphatase (H^+-PPase) :
H^+-PPase acidifies plant vacuoles or acidocalcisomes and maintains the pH gradient across the membrane. Our experiments revealed that a knockout mutant of H^+-PPase in Arabidopsis could not grow normally, suggesting that H^+-PPase is one of the essential membrane transport systems in plants.
(2) Biochemical properties and membrane topology of H^+-PPase :
In addition to plant H^+-PPase, we purified and characterized Streptomyces coelicolor H^+-PPase, since it and its mutant enzymes can be expressed in E. col. We revealed that S. coelicolor enzyme has a characteristic properties such as heat stability. We also determined the membrane topology of H^+-PPase and essential motifs in the enzyme. Our structural model has been accepted as the standard of H^+-PPases in various organisms.
(3) Crystallization of H^+-PPase :
We prepared a large quantity of Fv fragment, which specifically reacts with the active form of H+-PPase. We succeeded in preparation of a large quantity of the Fv-H+-PPase complex using the Fv fragment from the solubilized plant vacuolar membranes. Now, we are determining the best conditions for crystallization of the enzyme.
(4) Cation/H^+ exchangers :
Rice has five isoforms of cation/H^+ exchanger (CAX). These five isoforms varied with metal ion selectivity and tissue specific gene expression. CAXIa among them is expressed in all tissues examined and is highly expressed in companion and endodermis cells, which has high concentration of Ca^<2+>. We estimate that CAXla functions to reserve Ca^<2+> into vacuoles and is involved in the Ca^<2+> detoxification. The membrane topology and several functional motifs were also determined.
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[Journal Article] Disruption of AtPDR8, a plasma membrane ABC transporter of Arabidopsis, exhibits hypersensitive cell death by pathogen infection.2006
Author(s)
Kobae, Y., Yoshioka, H., Sekino, T., Nakanishi, Y., Martinoia, E., Maeshima, M.
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Journal Title
Plant Cell Physiol. 47
Pages: 309-318
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The involvement of tonoplast proton pumps and Na^+(K^+)/H^+ exchangers in the change of petal color during flower-opening of morning glory, Ipomoea tricolor cv. Heavenly Blue.2005
Author(s)
Yoshida, K., Kawachi, M., Mori, M., Maeshima, M., Kondo, M., Nishimura, M., Kondo, T.
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Journal Title
Plant Cell Physiol. 46
Pages: 407-415
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A new and simple technique for the isolation of plasma membrane lipids from root-tips.2005
Author(s)
Wagatsuma, T., Uemura, M., Mitsuhashi, W., Maeshima, M., Ishikawa, S., Kawamura, T., Murayama, T., Shiono, Y., Khan, M.S.H., Tawaraya, K.
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Journal Title
Soil Sci. Plant Nutr. 51
Pages: 135-139
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The distribution of aquaporins subtypes (PIP1, PIP2 and ^-TIP) is tissue dependent in soybean (Glycine max) root nodules.2005
Author(s)
Fleurat-Lessard, P., Michonneau, P., Maeshima, M., Drevon, J.J., Serraj, R.
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Journal Title
Annals of Botany 96
Pages: 457-460
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Effect of salt and osmotic stresses on the expression of genes for the vacuolar H^+-pyrophosphatase, H^+-ATPase subunit A and Na^+/H^+ antiporter from barley.2004
Author(s)
Fukuda, A., Chiba, K., Maeda, M., Nakamura, A.Maeshima, M., Tanaka, Y.
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Journal Title
J. Exp. Botany 55
Pages: 585-594
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Arabidopsis immunophilin-like TWD1 functionally interacts with vacuolar ABC transporters.2004
Author(s)
Geisler, M., Girin, M., Brnadt, S., Vincenzetti, V., Plaza, S., Paris, N., Kobae, Y., Maeshima, M.Billion, K., Kolukisaoglu, U.H., Schulz, B., Martinoia, E.
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Journal Title
Mol. Biol. Cell. 15
Pages: 3393-3405
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Isolation of intact vacuoles and proteomic analysis of tonoplast from suspension-cultured cells of Arabidopsis thaliana.2004
Author(s)
Shimaoka, T., Ohnishi, M., Sazuka, T., Mitsuhashi, N., Hara-Nishimura, I., Shimazaki, K., Maeshima, M., Yokota, A., Tomizawa, K., Mimura, T.
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Journal Title
Plant Cell Physiol. 45
Pages: 672-683
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Contribution of the plasma membrane and central vacuole in the formation of autolysosomes in tobacco cultured cells.2004
Author(s)
Yano, K., Matsui, S., Tsuchiya, T., Maeshima, M., Kutsuna, N., Hasezawa, S., Moriyasu, Y.
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Journal Title
Plant Cell Physiol. 45
Pages: 951-957
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Functional idientification of the glycerolaquaporin of Chlamydomonas reinhardtii CrMIP1.2004
Author(s)
Anderca, M.I., Suga, S., Furuich, T., Shimogawara, K., Maeshima, M., Muto S.
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Journal Title
Plant Cell Physiol. 45
Pages: 1313-1319
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis.2004
Author(s)
Kobae, Y., Uemura, T., Sato, M.H., Ohnishi, M., Mimura,.T., Maeshima, M.
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Journal Title
Plant Cell Physiol. 45
Pages: 1749-1758
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Functional expression of Acetabularia acetabulum vacuolar H+-pyrophosphatase in a yeast VMA3-deficient strain.2002
Author(s)
Ikeda, M., Umami, K., Hinohara, M., Tanimura, Y., Ohmae, A., Nakanishi, Y., Maeshima, M.
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Journal Title
J. Exp. Botany 53
Pages: 2273-2275
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Expression and distribution of vacuolar aquaporin in young and mature leaf tissues of Brassica napus in relation to water fluxes.2001
Author(s)
Frangne, N., Maeshima, M., Schaffner, A.R., Mandel, T., Martinoia E., Bonnemain, J.L.
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Journal Title
Planta 212
Pages: 270-278
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The protein storage vacuole : a unique compound organelle.2001
Author(s)
Jiang, L.Phillips, T.E., Hamm, C.A., Drozdowicz, Y.M., Rea, P.A., Maeshima, M., Rogers, S.W., Rogers, J.C.
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Journal Title
J. Cell Biol. 155
Pages: 991-1002
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Expression of V-ATPase proteolipid subunit of Acetabularia acetabulum in a VMA3-deficient strain of Saccharomyces cerevisiae and its complementation study.2001
Author(s)
Ikeda, M., Hinohara, M., Umami, K., Okada, Y., Wada, Y., Nakanishi, Y., Maeshima, M.
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Journal Title
Eur. J. Biochem. 268
Pages: 6097-6104
Description
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