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Purification of a tonoplast polypeptide with sucrose transport properties
Author(s) -
Thom Margaret,
Getz Hans Peter,
Maretzki Andrew
Publication year - 1992
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1992.tb01318.x
Subject(s) - sucrose , vacuole , biochemistry , monoclonal antibody , size exclusion chromatography , ion chromatography , chemistry , chromatography , transporter , biology , antibody , cytoplasm , enzyme , gene , immunology
Sucrose uptake into vacuoles of sugarcane parenchyma cells is mediated by a transporter. A polypeptide fitting that description has been partially purified from solubilized tonoplast by gel filtration and ion exchange chromatography. Purification of the relevant polypeptide was followed by reconstitution of fractions into liposomes and measurement of sucrose uptake into the proteoliposomes. In addition, a mono‐clonal antibody raised against tonoplast inhibited sucrose uptake. Western blots showed that the monoclonal antibody was relatively nonspecific but was suitable for monitoring the sucrose transport‐related polypeptide during purification. The mono‐clonal antibody bound a polypeptide of approximately 40 kDa, and this band was found in the fraction with sucrose uptake activity.