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

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Zeitschriftentitel:	The Scientific World Journal
Personen und Körperschaften:	De Caroli, Monica, Lenucci, Marcello S., Di Sansebastiano, Gian-Pietro, Tunno, Michela, Montefusco, Anna, Dalessandro, Giuseppe, Piro, Gabriella
In:	The Scientific World Journal, 2014, 2014, S. 1-7
Format:	E-Article
Sprache:	Englisch
veröffentlicht:	Hindawi Limited
Schlagwörter:	General Environmental Science General Biochemistry, Genetics and Molecular Biology General Medicine

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Zusammenfassung:	<jats:p><jats:italic>Cellulose synthase</jats:italic>-<jats:italic>like</jats:italic>(<jats:italic>Csl</jats:italic>) genes are believed to encode enzymes for the synthesis of cell wall matrix polysaccharides. The subfamily of<jats:italic>CslA</jats:italic>is putatively involved in the biosynthesis of<jats:italic>β</jats:italic>-mannans. Here we report a study on the cellular localization and the enzyme activity of an<jats:italic>Arabidopsis</jats:italic>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<jats:italic>in vitro</jats:italic>synthesis of a<jats:sup>14</jats:sup>C-mannan starting from GDP-<jats:sc>d</jats:sc>-[U-<jats:sup>14</jats:sup>C]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.</jats:p>
Umfang:	1-7
ISSN:	2356-6140 1537-744X
DOI:	10.1155/2014/792420