Isolation and Partial Characterization of Vacuoles from Tobacco Protoplasts 1
AUTOR(ES)
Mettler, Irvin J.
RESUMO
Protoplasts from suspension-cultured cells of Nicotiana glutinosa L. were lysed in 0.3 molar sorbitol in 2 millimolar ethylenediaminetetraacetate-tris(hydroxymethyl) aminomethane (pH 7.5) to release intact vacuoles. The vacuoles were purified by centrifugation in a Ficoll step gradient. About 11% of the vacuoles and 13% of the acid phosphatase activity was recovered in the purified vacuole fraction, suggesting that the vacuole is the major site for acid phosphatase in these cells. NADH-cytochrome c reductase, malate dehydrogenase, and cytochrome c oxidase activities were reduced during vacuole purification. The majority of the adenosine 5′-triphosphate (ATP) hydrolytic activity of purified vacuoles was associated with nonspecific acid phosphatase and not with a transport ATPase. As judged by acid phosphatase distribution and electron microscopy, the effective density of vacuoles in a sucrose gradient was low (less than 1.1 grams per cubic centimeter), although an unequivocal estimate of the vacuole or tonoplast density was not possible from the experiments conducted.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=543202Documentos Relacionados
- Vacuoles from Sugarcane Suspension Cultures 1: I. ISOLATION AND PARTIAL CHARACTERIZATION
- Isolation of Protoplasts and Vacuoles from Storage Tissue of Red Beet 1
- Isolation of Tonoplast Vesicles from Tobacco Protoplasts 1
- Isolation of Protoplasts and Vacuoles from Storage Tissue of Red Beet
- Isolation and Characterization of Vacuoles from Melilotus alba Mesophyll 1
