Partial purification of plant plasma membrane K+, Mg2+‐ATPase by ion exchange chromatography

A. Bérczi, D. James Morré

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

A purification procedure is presented which differs in three respects from other procedures for the purification of plant plasma membrane H+‐pumping ATPase (EC 3.6.1.35) from various plants. Soybean (Glycine max L. cv. Williams) hypocotyls were homogenized in the presence of physiological ionic strength and plasma membrane vesicles were purified by aqueous polymer two‐phase partitioning. Plasma membrane vesicles were then solubilized in one step by using non‐ionic detergent (either Triton X‐100 or C12E8). The Mg‐ATPase was separated by ion exchange chromatography from other solubilized membrane proteins. ATPase molecules bound to phosphocellulose fibers were eluted by a 0–1 M gradient of NaCl. The NaCl‐eluted fractions contained a Mg‐ATPase which showed the characteristics of Mg‐ATPase present in the plasma membranes. The specific activity of the partially purified enzyme was 2–5 μmol mg−1 min−1 when it was reconstituted into proteoliposomes. This value is in good agreement with data obtained by other purification methods in the literature. 1992 Physiologia Plantarum

Original languageEnglish
Pages (from-to)207-214
Number of pages8
JournalPhysiologia Plantarum
Volume85
Issue number2
DOIs
Publication statusPublished - 1992

Fingerprint

Ca(2+) Mg(2+)-ATPase
Ion Exchange Chromatography
ion exchange chromatography
adenosinetriphosphatase
plasma membrane
Cell Membrane
Soybeans
Hypocotyl
purification methods
Proton-Translocating ATPases
Octoxynol
ionic strength
membrane proteins
detergents
Detergents
Osmolar Concentration
hypocotyls
Adenosine Triphosphatases
Glycine max
polymers

Keywords

  • CE
  • Glycine max
  • ion exchange chromatography
  • K
  • Mg‐ATPase
  • non‐ionic detergent
  • phosphocellulose
  • plant plasma membrane
  • soybean
  • Triton X‐100

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science
  • Cell Biology

Cite this

Partial purification of plant plasma membrane K+, Mg2+‐ATPase by ion exchange chromatography. / Bérczi, A.; Morré, D. James.

In: Physiologia Plantarum, Vol. 85, No. 2, 1992, p. 207-214.

Research output: Contribution to journalArticle

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