Galactosyltransferases from Arabidopsis thaliana in the biosynthesis of type II arabinogalactan: molecular interaction enhances enzyme activity

Autor: Naomi Geshi, György Vereb, Alexander Schulz, Adiphol Dilokpimol, Satoshi Kaneko, Christian Peter Poulsen
Jazyk: angličtina
Rok vydání: 2014
Předmět:
Enzyme complex
Arabidopsis thaliana
Microsomes/metabolism
Arabidopsis
Golgi Apparatus
Plasma protein binding
Plant Science
Galactans/biosynthesis
Subcellular Fractions/enzymology
Galactans
Tobacco/metabolism
Protein O-glycosylation
chemistry.chemical_compound
Protein-protein interaction
Fluorescence Resonance Energy Transfer
Elméleti orvostudományok
Recombinant Proteins/metabolism
biology
Orvostudományok
Galactosyltransferases
Recombinant Proteins
Protein Transport
Biochemistry
Arabidopsis/enzymology
Biosynthetic process
Research Article
Subcellular Fractions
Protein Binding
Glycan
Glycosylation
Green Fluorescent Proteins
Galactosyltransferases/metabolism
Arabinogalactan
Microsomes
Tobacco
Galactose/metabolism
Golgi Apparatus/enzymology
Plant cell wall
Arabinogalactan protein
Green Fluorescent Proteins/metabolism
Galactosyltransferase
Arabidopsis Proteins
Galactose
Plant Leaves
chemistry
FRET
biology.protein
Arabidopsis Proteins/metabolism
Plant Leaves/metabolism
Zdroj: Dilokpimol, A, Poulsen, C P, Vereb, G, Kaneko, S, Schulz, A & Geshi, N 2014, ' Galactosyltransferases from Arabidopsis thaliana in the biosynthesis of type II arabinogalactan: molecular interaction enhances enzyme activity ', B M C Plant Biology, vol. 14, 90 . https://doi.org/10.1186/1471-2229-14-90
Dilokpimol, A, Poulsen, C P, Vereb, G, Kaneko, S, Schulz, A & Geshi, N 2014, ' Galactosyltransferases from Arabidopsis thaliana in the biosynthesis of type II arabinogalactan : molecular interaction enhances enzyme activity ', BMC Plant Biology, vol. 14, pp. 90 . https://doi.org/10.1186/1471-2229-14-90
BMC Plant Biology
DOI: 10.1186/1471-2229-14-90
Popis: BACKGROUND: Arabinogalactan proteins are abundant proteoglycans present on cell surfaces of plants and involved in many cellular processes, including somatic embryogenesis, cell-cell communication and cell elongation. Arabinogalactan proteins consist mainly of glycan, which is synthesized by post- translational modification of proteins in the secretory pathway. Importance of the variations in the glycan moiety of arabinogalactan proteins for their functions has been implicated, but its biosynthetic process is poorly understood.\n\nRESULTS: We have identified a novel enzyme in the biosynthesis of the glycan moiety of arabinogalactan proteins. The At1g08280 (AtGALT29A) from Arabidopsis thaliana encodes a putative glycosyltransferase (GT), which belongs to the Carbohydrate Active Enzyme family GT29. AtGALT29A co-expresses with other arabinogalactan GTs, AtGALT31A and AtGLCAT14A. The recombinant AtGALT29A expressed in Nicotiana benthamiana demonstrated a galactosyltransferase activity, transferring galactose from UDP-galactose to a mixture of various oligosaccharides derived from arabinogalactan proteins. The galactose-incorporated products were analyzed using structure-specific hydrolases indicating that the recombinant AtGALT29A possesses beta-1,6-galactosyltransferase activity, elongating beta-1,6-galactan side chains and forming 6-Gal branches on the beta-1,3-galactan main chain of arabinogalactan proteins. The fluorescence tagged AtGALT29A expressed in N. benthamiana was localized to Golgi stacks where it interacted with AtGALT31A as indicated by Forster resonance energy transfer. Biochemically, the enzyme complex containing AtGALT31A and AtGALT29A could be co-immunoprecipitated and the isolated protein complex exhibited increased level of beta-1,6-galactosyltransferase activities compared to AtGALT29A alone.\n\nCONCLUSIONS: AtGALT29A is a beta-1,6-galactosyltransferase and can interact with AtGALT31A. The complex can work cooperatively to enhance the activities of adding galactose residues 6-linked to beta-1,6-galactan and to beta-1,3-galactan. The results provide new knowledge of the glycosylation process of arabinogalactan proteins and the functional significance of protein-protein interactions among O-glycosylation enzymes.
Databáze: OpenAIRE
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