Dimer formation of subunit G of the yeast V-ATPase

Andrea Armbrüster; Susanne M. Bailer; Michel H.J. Koch; Jasminka Godovac-Zimmermann; Gerhard Grüber

doi:10.1016/S0014-5793(03)00643-4

Dimer formation of subunit G of the yeast V-ATPase

Andrea Armbrüster, Susanne M. Bailer, Michel H.J. Koch, Jasminka Godovac-Zimmermann, Gerhard Grüber^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

17 Citations (Scopus)

Abstract

The G subunit of the vacuolar ATPase (V-ATPase) is a component of the stalk connecting the V₁ and V_O sectors of the enzyme and is essential for normal assembly and function. Subunit G (Vma10p) of the yeast V-ATPase was expressed in Escherichia coli as a soluble protein and was purified to homogeneity. The molecular mass of subunit G, determined by Native-polyacrylamide gel electrophoresis, gel filtration analysis and small-angle X-ray scattering, was approximately 28±2 kDa, indicating that this protein is dimeric. With a radius of gyration (R_g) and a maximum size (D_max) of 2.7±0.2 nm and 8.0±0.3 nm, respectively, the G-dimer is rather elongated. To understand which region of subunit G is required to mediate dimerization, a G_38-144 form (the carboxyl-terminus) was expressed and purified. G_38-144 is homogeneous, with a molecular mass of approximately 12±3 kDa, indicating a monomeric form in solution.

Original language	English
Pages (from-to)	395-400
Number of pages	6
Journal	FEBS Letters
Volume	546
Issue number	2-3
DOIs	https://doi.org/10.1016/S0014-5793(03)00643-4
Publication status	Published - Jul 10 2003
Externally published	Yes

ASJC Scopus Subject Areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Keywords

Dimerization
Small-angle X-ray scattering
V ATPase
VV ATPase
Vacuolar-type ATPase
Vma10p

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10.1016/S0014-5793(03)00643-4

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title = "Dimer formation of subunit G of the yeast V-ATPase",

abstract = "The G subunit of the vacuolar ATPase (V-ATPase) is a component of the stalk connecting the V1 and VO sectors of the enzyme and is essential for normal assembly and function. Subunit G (Vma10p) of the yeast V-ATPase was expressed in Escherichia coli as a soluble protein and was purified to homogeneity. The molecular mass of subunit G, determined by Native-polyacrylamide gel electrophoresis, gel filtration analysis and small-angle X-ray scattering, was approximately 28±2 kDa, indicating that this protein is dimeric. With a radius of gyration (Rg) and a maximum size (Dmax) of 2.7±0.2 nm and 8.0±0.3 nm, respectively, the G-dimer is rather elongated. To understand which region of subunit G is required to mediate dimerization, a G38-144 form (the carboxyl-terminus) was expressed and purified. G38-144 is homogeneous, with a molecular mass of approximately 12±3 kDa, indicating a monomeric form in solution.",

keywords = "Dimerization, Small-angle X-ray scattering, V ATPase, VV ATPase, Vacuolar-type ATPase, Vma10p",

author = "Andrea Armbr{\"u}ster and Bailer, {Susanne M.} and Koch, {Michel H.J.} and Jasminka Godovac-Zimmermann and Gerhard Gr{\"u}ber",

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T1 - Dimer formation of subunit G of the yeast V-ATPase

AU - Armbrüster, Andrea

AU - Bailer, Susanne M.

AU - Koch, Michel H.J.

AU - Godovac-Zimmermann, Jasminka

AU - Grüber, Gerhard

PY - 2003/7/10

Y1 - 2003/7/10

N2 - The G subunit of the vacuolar ATPase (V-ATPase) is a component of the stalk connecting the V1 and VO sectors of the enzyme and is essential for normal assembly and function. Subunit G (Vma10p) of the yeast V-ATPase was expressed in Escherichia coli as a soluble protein and was purified to homogeneity. The molecular mass of subunit G, determined by Native-polyacrylamide gel electrophoresis, gel filtration analysis and small-angle X-ray scattering, was approximately 28±2 kDa, indicating that this protein is dimeric. With a radius of gyration (Rg) and a maximum size (Dmax) of 2.7±0.2 nm and 8.0±0.3 nm, respectively, the G-dimer is rather elongated. To understand which region of subunit G is required to mediate dimerization, a G38-144 form (the carboxyl-terminus) was expressed and purified. G38-144 is homogeneous, with a molecular mass of approximately 12±3 kDa, indicating a monomeric form in solution.

AB - The G subunit of the vacuolar ATPase (V-ATPase) is a component of the stalk connecting the V1 and VO sectors of the enzyme and is essential for normal assembly and function. Subunit G (Vma10p) of the yeast V-ATPase was expressed in Escherichia coli as a soluble protein and was purified to homogeneity. The molecular mass of subunit G, determined by Native-polyacrylamide gel electrophoresis, gel filtration analysis and small-angle X-ray scattering, was approximately 28±2 kDa, indicating that this protein is dimeric. With a radius of gyration (Rg) and a maximum size (Dmax) of 2.7±0.2 nm and 8.0±0.3 nm, respectively, the G-dimer is rather elongated. To understand which region of subunit G is required to mediate dimerization, a G38-144 form (the carboxyl-terminus) was expressed and purified. G38-144 is homogeneous, with a molecular mass of approximately 12±3 kDa, indicating a monomeric form in solution.

KW - Dimerization

KW - Small-angle X-ray scattering

KW - V ATPase

KW - VV ATPase

KW - Vacuolar-type ATPase

KW - Vma10p

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ER -

Dimer formation of subunit G of the yeast V-ATPase

Abstract

ASJC Scopus Subject Areas

Keywords

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