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Calmodulin-stimulated Ca2+-ATPases in the vacuolar and plasma membranes in cauliflower
Högskolan i Jönköping, Högskolan för lärande och kommunikation, HLK, Ämnesforskning. Department of Plant Biochemistry, Lund University, Lund, Sweden.ORCID-id: 0000-0003-0117-2974
1997 (Engelska)Ingår i: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 114, nr 3, s. 999-1007Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The subcellular locations of Ca2+-ATPases in the membranes of cauliflower (Brassica oleracea L.) inflorescences were investigated. After continuous sucrose gradient centrifugation a 111-kD calmodulin (CaM)-stimulated and CaM-binding Ca2+-ATPase (BCA1; P. Askerlund [1996] Plant Physiol 110: 913–922; S. Malmstrom, P. Askerlund, M.G. Palmgren [1997] FEBS Lett 400: 324–328) comigrated with vacuolar membrane markers, whereas a 116-kD CaM-binding Ca2+-ATPase co-migrated with a marker for the plasma membrane. The 116-kD Ca2+-ATPase was enriched in plasma membranes obtained by aqueous two-phase partitioning, which is in agreement with a plasma membrane location of this Ca2+-ATPase. Countercurrent distribution of a low-density intracellular membrane fraction in an aqueous two-phase system resulted in the separation of the endoplasmic reticulum and vacuolar membranes. The 111-kD Ca2+-ATPase co-migrated with a vacuolar membrane marker after countercurrent distribution but not with markers for the endoplasmic reticulum. A vacuolar membrane location of the 111-kD Ca2+-ATPase was further supported by experiments with isolated vacuoles from cauliflower: (a) Immunoblotting with an antibody against the 111-kD Ca2+-ATPase showed that it was associated with the vacuoles, and (b) ATP-dependent Ca2+ uptake by the intact vacuoles was found to be CaM stimulated and partly protonophore insensitive.

Ort, förlag, år, upplaga, sidor
1997. Vol. 114, nr 3, s. 999-1007
Nyckelord [en]
Calcium-Transporting ATPases/isolation & purification/*metabolism, Calmodulin/*pharmacology, Cell Fractionation, Cell Membrane/enzymology/ultrastructure, Centrifugation; Density Gradient, Egtazic Acid/pharmacology, Vacuoles/*enzymology/ultrastructure, Vegetables/*enzymology
Nationell ämneskategori
Biokemi och molekylärbiologi
Identifikatorer
URN: urn:nbn:se:hj:diva-2173PubMedID: 9232880OAI: oai:DiVA.org:hj-2173DiVA, id: diva2:32993
Tillgänglig från: 2007-10-17 Skapad: 2007-10-17 Senast uppdaterad: 2017-12-12Bibliografiskt granskad

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