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Journal of Experimental Botany, Vol. 54, No. 386, pp. 1407-1414, May 1, 2003
© 2003 Oxford University Press

Evidence for a tonoplast-associated form of sucrose synthase and its potential involvement in sucrose mobilization from the vacuole

Received 11 November 2002; Accepted 3 February 2003

Ed Etxeberria1, and Pedro Gonzalez

Citrus Research and Education Center, IFAS, University of Florida, 700 Experiment Station Road, Lake Alfred, Fl 33850-2299, USA

1 To whom correspondence should be addressed. Fax: +1 863 956 4631. E-mail: eje{at}lal.ufl.edu

The following work presents new evidence for a tonoplast localization of sucrose synthase and its functional role during the mobilization of vacuolar sucrose. Highly purified tonoplast vesicles were associated with significant SuSy levels as determined by immuno-recognition, enzymatic activity, and by-product measurements. Total tonoplast-bound SuSy was estimated to be approximately 7% of the total tissue activity. SuSy affinity to the tonoplast was confirmed by the lack of SuSy displacement by ionic washes and also by the tonoplast ability to bind to exogenously added SuSy as compared to the cytosolic marker alcohol dehydrogenase. UL-[14C]sucrose-loaded vesicles incubated with ATP and UDP produced [14C]UDP-Glc as determined by UDP-Glc dehydrogenase and by the ability of the product to bind to DEAE-cellulose and to co-migrate with authentic UDP-Glc on TLC. ATP alone induced sucrose efflux but not the production of [14C]ADP-Glc. Kinetic analysis of [14C]UDPG formation under conditions of low sucrose availability suggests sucrose channelling between the ATP-dependent sucrose transporter and SuSy, thus corroborating the association of SuSy with the tonoplast and its involvement in sucrose mobilization from the vacuole.

Key words: Red beet, sucrose efflux, sucrose mobilization, sucrose synthase, tonoplast transport.


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