Purification and functional reconstitution of the tonoplast pyrophosphate-dependent proton pump from Rubus hispidus cell cultures

Détails

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Etat: Public
Version: de l'auteur
ID Serval
serval:BIB_1CC0403A9DC2
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Purification and functional reconstitution of the tonoplast pyrophosphate-dependent proton pump from Rubus hispidus cell cultures
Périodique
Plant Science
Auteur(s)
Perotti E., Gavin O., Widmer F., Chanson A.
ISSN
0168-9452
Statut éditorial
Publié
Date de publication
1994
Peer-reviewed
Oui
Volume
103
Numéro
1
Pages
25-31
Langue
anglais
Résumé
The hydrolytic subunit of the H+-translocating inorganic pyrophosphatase (V-PPase EC 3.6.1.1.) prepared from Rubus hispidus cell cultures has been purified from tonoplast-enriched membranes and analysed by SDS-polyacrylamide gel electrophoresis, Only one polypeptide of M(r) 70 000 was recovered with the V-PPase activity after solubilization in the presence of Triton X-100, purification by gel filtration (Superose) and anion exchange (Mono Q) chromatography. This polypeptide strongly cross-reacted with an antibody raised against the V-PPase from Vigna radiata. The tonoplast-enriched fraction was also used to solubilize and reconstitute the-V-PPase. The proteoliposomes showing a PPi-dependent proton transport activity were purified by gel filtration (Superose) and analysed by SDS-polyacrylamide gel electrophoresis. Only one polypeptide of M(r) 70 000 was recovered with the proton-pumping activity. All these data suggest that the native V-PPase from Rubus is composed of a single kind of polypeptide with an M(r) of 70 000 and representing the catalytic subunit.
Mots-clé
PYROPHOSPHATASE, PROTON PUMP, PURIFICATION, SOLUBILIZATION AND RECONSTITUTION, TONOPLAST, RUBUS HISPIDUS, TRANSLOCATING INORGANIC PYROPHOSPHATASE, SUBSTRATE-BINDING SUBUNIT, VACUOLAR, TRANSPORT
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Création de la notice
13/08/2015 8:01
Dernière modification de la notice
20/08/2019 12:53
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