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Approaching determination of H+-ATPase H+/ATP stoichiometry

Wosicki, Stanislaw LU (2011) KEMR13 20111
Department of Chemistry
Abstract
The plant plasma membrane H+-ATPase was activated by the binding of a 14-3-3 protein. This complex can be stabilized by the fungal toxin - fusicoccin. During this activation H+/ATP stoichiometry changed (1). This work presents approaching to find the new stoichiometry. The main part presents looking for the best preparation of plasma membrane vesicles for measurements. A new pH indicator – Glu3 – was trapped inside vesicles in a few different ways such as Brij 58 treatment or sonication. Membranes were washed by ultracentrifugations or gel chromatography. The tight of prepared vesicles was examined by acid titration. The best result was achieved by sonication and gel chromatography. Brij 58 also affects the size of plasma membrane vesicles... (More)
The plant plasma membrane H+-ATPase was activated by the binding of a 14-3-3 protein. This complex can be stabilized by the fungal toxin - fusicoccin. During this activation H+/ATP stoichiometry changed (1). This work presents approaching to find the new stoichiometry. The main part presents looking for the best preparation of plasma membrane vesicles for measurements. A new pH indicator – Glu3 – was trapped inside vesicles in a few different ways such as Brij 58 treatment or sonication. Membranes were washed by ultracentrifugations or gel chromatography. The tight of prepared vesicles was examined by acid titration. The best result was achieved by sonication and gel chromatography. Brij 58 also affects the size of plasma membrane vesicles what was demonstrated. The average diameter decreases with increasing concentration of detergent. (Less)
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author
Wosicki, Stanislaw LU
supervisor
organization
course
KEMR13 20111
year
type
H2 - Master's Degree (Two Years)
subject
keywords
biokemi
language
English
id
3053555
date added to LUP
2012-09-19 11:28:30
date last changed
2012-09-19 11:28:30
@misc{3053555,
  abstract     = {The plant plasma membrane H+-ATPase was activated by the binding of a 14-3-3 protein. This complex can be stabilized by the fungal toxin - fusicoccin. During this activation H+/ATP stoichiometry changed (1). This work presents approaching to find the new stoichiometry. The main part presents looking for the best preparation of plasma membrane vesicles for measurements. A new pH indicator – Glu3 – was trapped inside vesicles in a few different ways such as Brij 58 treatment or sonication. Membranes were washed by ultracentrifugations or gel chromatography. The tight of prepared vesicles was examined by acid titration. The best result was achieved by sonication and gel chromatography. Brij 58 also affects the size of plasma membrane vesicles what was demonstrated. The average diameter decreases with increasing concentration of detergent.},
  author       = {Wosicki, Stanislaw},
  keyword      = {biokemi},
  language     = {eng},
  note         = {Student Paper},
  title        = {Approaching determination of H+-ATPase H+/ATP stoichiometry},
  year         = {2011},
}