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Identification and localization of calreticulin in plant cells
Authors:M Opas  S Tharin  R E Milner  M Michalak
Institution:(1) Department of Anatomy and Cell Biology, Medical Sciences Building, University of Toronto, M5S 1A8 Toronto, Ont., Canada;(2) MRC Group in Molecular Biology of Membrane, Department of Biochemistry, University of Alberta, Edmonton, Alberta
Abstract:Summary In the present study we have investigated the presence and distribution of calreticulin in plant protoplasts. Calreticulin was purified from plant homogenates using a selective ammonium sulfate precipitation procedure developed for the purification of mammalian calreticulins and shown to bind calcium in45Ca2+ overlay assays. The protein was localized to plant cell endoplasmic reticulum by the indirect immunofluorescence staining of protoplasts with anti-calreticulin antibodies. No calreticulin was observed within large vacuoles. We conclude that calreticulin is present in the endoplasmic reticulum of plant cells, where, by analogy to the mammalian endoplasmic reticulum, it may play a major role in Ca2+ binding and storage.Abbreviations ER endoplasmic reticulum - SR sarcoplasmic reticulum - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - PBS phosphate-buffered saline
Keywords:Calcium binding proteins  Calreticulin  Endoplasmic reticulum  Plants
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