Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum. / Villalba, J. M.; Palmgren, M. G.; Berberian, G. E.; Ferguson, C.; Serrano, R.

In: Journal of Biological Chemistry, Vol. 267, No. 17, 1992, p. 12341-12349.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Villalba, JM, Palmgren, MG, Berberian, GE, Ferguson, C & Serrano, R 1992, 'Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum', Journal of Biological Chemistry, vol. 267, no. 17, pp. 12341-12349.

APA

Villalba, J. M., Palmgren, M. G., Berberian, G. E., Ferguson, C., & Serrano, R. (1992). Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum. Journal of Biological Chemistry, 267(17), 12341-12349.

Vancouver

Villalba JM, Palmgren MG, Berberian GE, Ferguson C, Serrano R. Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum. Journal of Biological Chemistry. 1992;267(17):12341-12349.

Author

Villalba, J. M. ; Palmgren, M. G. ; Berberian, G. E. ; Ferguson, C. ; Serrano, R. / Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 17. pp. 12341-12349.

Bibtex

@article{a674b5ac0147441082b87e56314a05a4,
title = "Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum",
abstract = "Recombinant plant plasma membrane H+-ATPase has been produced in a yeast expression system comprising a multicopy plasmid and the strong promoter of the yeast PMA1 gene. Western blotting with a specific monoclonal antibody showed that the plant ATPase is one of the major membrane proteins made by the transformed cells, accounting for about 1% of total yeast protein. The plant ATPase synthesized in yeast is fully active. It hydrolyzes ATP, pumps protons, and the reaction cycle involves a phosphorylated intermediate. Phosphorylation is possible from both ATP and P1. Unlike the situation in plants, however, most of the plant ATPase is not expressed in the yeast plasma membrane. Rather, the enzyme appears to remain trapped at a very early stage of secretory pathway: insertion into the endoplasmic reticulum. This organelle was observed to proliferate in the form of stacked membranes surrounding the yeast nucleus in order to accommodate the large amount of plant ATPase produced. In this location, the plant ATPase can be purified with high yield (70 mg from 1 kg of yeast) from membranes devoid of endogenous yeast plasma membrane H+-ATPase. This convenient expression system could be useful for other eukaryotic membrane proteins and ATPases.",
author = "Villalba, {J. M.} and Palmgren, {M. G.} and Berberian, {G. E.} and C. Ferguson and R. Serrano",
year = "1992",
language = "English",
volume = "267",
pages = "12341--12349",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "17",

}

RIS

TY - JOUR

T1 - Functional expression of plant plasma membrane H+-ATPase in yeast endoplasmic reticulum

AU - Villalba, J. M.

AU - Palmgren, M. G.

AU - Berberian, G. E.

AU - Ferguson, C.

AU - Serrano, R.

PY - 1992

Y1 - 1992

N2 - Recombinant plant plasma membrane H+-ATPase has been produced in a yeast expression system comprising a multicopy plasmid and the strong promoter of the yeast PMA1 gene. Western blotting with a specific monoclonal antibody showed that the plant ATPase is one of the major membrane proteins made by the transformed cells, accounting for about 1% of total yeast protein. The plant ATPase synthesized in yeast is fully active. It hydrolyzes ATP, pumps protons, and the reaction cycle involves a phosphorylated intermediate. Phosphorylation is possible from both ATP and P1. Unlike the situation in plants, however, most of the plant ATPase is not expressed in the yeast plasma membrane. Rather, the enzyme appears to remain trapped at a very early stage of secretory pathway: insertion into the endoplasmic reticulum. This organelle was observed to proliferate in the form of stacked membranes surrounding the yeast nucleus in order to accommodate the large amount of plant ATPase produced. In this location, the plant ATPase can be purified with high yield (70 mg from 1 kg of yeast) from membranes devoid of endogenous yeast plasma membrane H+-ATPase. This convenient expression system could be useful for other eukaryotic membrane proteins and ATPases.

AB - Recombinant plant plasma membrane H+-ATPase has been produced in a yeast expression system comprising a multicopy plasmid and the strong promoter of the yeast PMA1 gene. Western blotting with a specific monoclonal antibody showed that the plant ATPase is one of the major membrane proteins made by the transformed cells, accounting for about 1% of total yeast protein. The plant ATPase synthesized in yeast is fully active. It hydrolyzes ATP, pumps protons, and the reaction cycle involves a phosphorylated intermediate. Phosphorylation is possible from both ATP and P1. Unlike the situation in plants, however, most of the plant ATPase is not expressed in the yeast plasma membrane. Rather, the enzyme appears to remain trapped at a very early stage of secretory pathway: insertion into the endoplasmic reticulum. This organelle was observed to proliferate in the form of stacked membranes surrounding the yeast nucleus in order to accommodate the large amount of plant ATPase produced. In this location, the plant ATPase can be purified with high yield (70 mg from 1 kg of yeast) from membranes devoid of endogenous yeast plasma membrane H+-ATPase. This convenient expression system could be useful for other eukaryotic membrane proteins and ATPases.

UR - http://www.scopus.com/inward/record.url?scp=0026653810&partnerID=8YFLogxK

M3 - Journal article

C2 - 1534807

AN - SCOPUS:0026653810

VL - 267

SP - 12341

EP - 12349

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 17

ER -

ID: 245002327