title: Cellular localization and biochemical characterization of a chimeric fluorescent protein fusion of Arabidopsis cellulose synthase-like A2 inserted into Golgi membrane.
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.
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Monica
authors.identifier: 0000-0001-8668-4711
authors.initials: M
authors.lastname: De Caroli
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Marcello S
authors.initials: MS
authors.lastname: Lenucci
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Gian-Pietro
authors.identifier: 0000-0001-5388-0695
authors.initials: GP
authors.lastname: Di Sansebastiano
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Michela
authors.identifier: 0000-0003-2452-7598
authors.initials: M
authors.lastname: Tunno
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Montefusco
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Giuseppe
authors.identifier: 0000-0003-0820-4000
authors.initials: G
authors.lastname: Dalessandro
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento (DiSTeBA), Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Gabriella
authors.identifier: 0000-0002-3420-8788
authors.initials: G
authors.lastname: Piro
mesh_terms: D017360:Arabidopsis / Q000201:enzymology; D001669:Biochemical Phenomena; D002678:Chimera / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry* / Q000378:metabolism; D018515:Plant Leaves / Q000201:enzymology; D014026:Nicotiana / Q000201:enzymology; D021601:trans-Golgi Network / Q000737:chemistry* / Q000201:enzymology
chemical_list: D005456:Fluorescent Dyes; D005964:Glucosyltransferases; C478648:cellulose synthase