Journal Article

A 64 kDa sucrose binding protein is membrane-associated and tonoplast-localized in developing mung bean seeds

Junqi Wang, Pui Kit Suen, Zeng-Fu Xu and Liwen Jiang

in Journal of Experimental Botany

Published on behalf of Society for Experimental Biology

Volume 60, issue 2, pages 629-639
Published in print February 2009 | ISSN: 0022-0957
Published online January 2009 | e-ISSN: 1460-2431 | DOI: https://dx.doi.org/10.1093/jxb/ern308

More Like This

Show all results sharing this subject:

  • Plant Sciences and Forestry

GO

Show Summary Details

Preview

Sucrose binding proteins (SBPs) were predicted to be membrane-associated, but have been shown to localize in the lumen of protein storage vacuoles of various seeds. In this study, a new 64 kDa SBP has been identified from developing mung bean (Vigna radiata) seeds (here termed VrSBP1) via MS/MS analysis and N-terminal amino acid sequencing analysis and specific antibodies were generated using purified VrSBP1 proteins. Western blot analysis with the new VrSBP1 antibodies showed that, similar to most seed storage proteins, VrSBP1 proteins accumulated during seed development and were subsequently mobilized once the mung bean seeds germinated. Immunogold electron microscope (EM) studies on ultra-thin sections of high-pressure freezing/frozen substituted developing mung bean cotyledons demonstrated that VrSBP1 was localized specifically to the tonoplast of the protein storage vacuole and to the limiting membrane of a novel putative prevacuolar compartment. Biochemical and subcellular fractionation studies further demonstrated that VrSBP1 proteins were membrane-associated in developing mung beans, consistent with their tonoplast localization. This study thus shows convincing evidence of tonoplast-localization of a plant SBP for its future functional characterization and provides a model of studying non-integral membrane proteins associated with the tonoplasts in plant cells.

Keywords: High-pressure freezing; immunogold EM; membrane-associated; mung bean; sucrose binding proteins; tonoplast

Journal Article.  5560 words.  Illustrated.

Subjects: Plant Sciences and Forestry