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The Scientific World Journal
Volume 2014, Article ID 792420, 7 pages
http://dx.doi.org/10.1155/2014/792420
Research Article

Cellular Localization and Biochemical Characterization of a Chimeric Fluorescent Protein Fusion of Arabidopsis Cellulose Synthase-Like A2 Inserted into Golgi Membrane

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy

Received 29 August 2013; Accepted 27 October 2013; Published 14 January 2014

Academic Editors: L. Costa, D. Sarkar, B. Vyskot, and C. R. Wilson

Copyright © 2014 Monica De Caroli et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Cellulose synthase-like (Csl) genes are believed to encode enzymes for the synthesis of cell wall matrix polysaccharides. The subfamily of CslA is putatively involved in the biosynthesis of β-mannans. Here we report a study on the cellular localization and the enzyme activity of an Arabidopsis CslA family member, AtCslA2. We show that the fluorescent protein fusion AtCslA2-GFP, transiently expressed in tobacco leaf protoplasts, is synthesized in the ER and it accumulates in the Golgi stacks. The chimera is inserted in the Golgi membrane and is functional since membrane preparations obtained by transformed protoplasts carry out the in vitro synthesis of a 14C-mannan starting from GDP-d-[U-14C]mannose as substrate. The enzyme specific activity is increased by approximately 38% in the transformed protoplasts with respect to wild-type. Preliminary tests with proteinase K, biochemical data, and TM domain predictions suggest that the catalytic site of AtCslA2 faces the Golgi lumen.