Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis

Dietz K-J, Rudloff S, Ageorges A, Eckerskorn C, Fischer K, Arbinger B (1995)
The Plant Journal 8(4): 521-529.

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Abstract
A tonoplast protein of 31 kDa apparent molecular mass (TpP 31) was isolated from two-dimensional gels. Amino acid sequences were determined from LysC endoproteinase-peptide fragments. Using degenerate oligonucleotides, a corresponding cDNA clone of 1034 bp was isolated from a barley leaf cDNA library. It encodes for subunit E of the vacuolar H+-ATPase, the first one identi fled in plants so far. The open reading frame extends over 681 bp, encoding a gene product of 227 amino acids and a calculated molecular weight of 26 228 g mol(-1). Northern and Western blot analysis indicates constitutive expression of subunit E in all plant organs with only small effects of salt stress. Localization of TpP 31 at the tonoplast was confirmed in fractions of purified vacuolar membrane obtained by free-flow electrophoresis. Immunoprecipitation of newly synthesized S-35-labelled membrane proteins with anti-TpP 31 gave two additional bands with apparent molecular masses of about 53 and 62 kDa. Gel filtration after mild solubilization showed co-purification of TpP 31 with the 55 kDa subunit of the H+-ATPase. Both results provide evidence beyond the sequence homology that TpP 31 is a structural component of the vacuolar H+-ATPase.
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Dietz K-J, Rudloff S, Ageorges A, Eckerskorn C, Fischer K, Arbinger B. Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis. The Plant Journal. 1995;8(4):521-529.
Dietz, K. - J., Rudloff, S., Ageorges, A., Eckerskorn, C., Fischer, K., & Arbinger, B. (1995). Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis. The Plant Journal, 8(4), 521-529.
Dietz, K. - J., Rudloff, S., Ageorges, A., Eckerskorn, C., Fischer, K., and Arbinger, B. (1995). Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis. The Plant Journal 8, 521-529.
Dietz, K.-J., et al., 1995. Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis. The Plant Journal, 8(4), p 521-529.
K.-J. Dietz, et al., “Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis”, The Plant Journal, vol. 8, 1995, pp. 521-529.
Dietz, K.-J., Rudloff, S., Ageorges, A., Eckerskorn, C., Fischer, K., Arbinger, B.: Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis. The Plant Journal. 8, 521-529 (1995).
Dietz, Karl-Josef, Rudloff, S., Ageorges, A., Eckerskorn, C., Fischer, K., and Arbinger, B. “Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis”. The Plant Journal 8.4 (1995): 521-529.
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