Search

Your search keyword '"Yi Hsiang Chou"' showing total 4 results
Start Over Author "Yi Hsiang Chou" Topic arabidopsis thaliana
4 results on '"Yi Hsiang Chou"'

1. Protein-protein interactions among xyloglucan-synthesizing enzymes and formation of Golgi-localized multiprotein complexes

Author

Olga A. Zabotina, Gennady Pogorelko, Yi-Hsiang Chou, and Zachary T. Young

Subjects
Multiprotein complex, Fucosyltransferase, Physiology, Arabidopsis, Golgi Apparatus, Plant Science, Fluorescence, Protein–protein interaction, Bimolecular fluorescence complementation, chemistry.chemical_compound, symbols.namesake, Catalytic Domain, Protein Interaction Mapping, Arabidopsis thaliana, Immunoprecipitation, Glucans, biology, Chemistry, Arabidopsis Proteins, Protoplasts, Cell Biology, General Medicine, Golgi apparatus, biology.organism_classification, Galactosyltransferases, Xyloglucan, Protein Subunits, Biochemistry, Multiprotein Complexes, biology.protein, symbols, Xylans, Protein Binding
Abstract

Arabidopsis thaliana xyloglucan has an XXXG structure, with branches of xylosyl residues, β-D-galacosyl-(1,2)-α-d-xylosyl motifs and fucosylated β-D-galactosyl-(1,2)-α-D-xylosyl motifs. Most of the enzymes involved in xyloglucan biosynthesis in Arabidopsis have been identified, including the glucan synthase CSLC4 (cellulose synthase-like C4), three xylosyltransferases (XXT1, XXT2 and XXT5), two galactosyltransferases (MUR3 and XLT2) and the fucosyltransferase FUT1. The XXTs and CSLC4 form homo- and heterocomplexes and were proposed to co-localize in the same complex, but the organization of the other xyloglucan-synthesizing enzymes remains unclear. Here we investigate whether the glycosyltransferases MUR3, XLT2 and FUT1 interact with the XXT-CSLC4 complexes in the Arabidopsis Golgi. We used co-immunoprecipitation and bimolecular fluorescence complementation, with signal quantification by flow cytometry, to demonstrate that CSLC4 interacts with MUR3, XLT2 and FUT1. FUT1 forms homocomplexes and interacts with MUR3, XLT2, XXT2 and XXT5. XLT2 interacts with XXT2 and XXT5, but MUR3 does not. Co-immunoprecipitation assays showed that FUT1 forms a homocomplex through disulfide bonds, and formation of the heterocomplexes does not involve covalent interactions. In vitro pull-down assays indicated that interactions in the FUT1-MUR3 and FUT1-XXT2 complexes occur through the protein catalytic domains. We propose that enzymes involved in xyloglucan biosynthesis are functionally organized in multiprotein complexes localized in the Golgi.

Published
2014

2. Xyloglucan xylosyltransferases XXT1, XXT2, and XXT5 and the glucan synthase CSLC4 form Golgi-localized multiprotein complexes

Author

Olga A. Zabotina, Yi-Hsiang Chou, and Gennady Pogorelko

Subjects
Multiprotein complex, Physiology, Recombinant Fusion Proteins, Immunoblotting, Arabidopsis, Biochemical Processes and Macromolecular Structures, Golgi Apparatus, Plant Science, Cell wall, chemistry.chemical_compound, symbols.namesake, Bimolecular fluorescence complementation, Genetics, Arabidopsis thaliana, Immunoprecipitation, Pentosyltransferases, Golgi membrane, biology, Arabidopsis Proteins, Protoplasts, Golgi apparatus, biology.organism_classification, Flow Cytometry, Xyloglucan, Protein Transport, chemistry, Biochemistry, Microscopy, Fluorescence, Glucosyltransferases, Multiprotein Complexes, symbols, Protein Multimerization, Plasmids, Protein Binding
Abstract

Xyloglucan is the major hemicellulosic polysaccharide in the primary cell walls of most vascular dicotyledonous plants and has important structural and physiological functions in plant growth and development. In Arabidopsis (Arabidopsis thaliana), the 1,4-β-glucan synthase, Cellulose Synthase-Like C4 (CSLC4), and three xylosyltransferases, XXT1, XXT2, and XXT5, act in the Golgi to form the xylosylated glucan backbone during xyloglucan biosynthesis. However, the functional organization of these enzymes in the Golgi membrane is currently unknown. In this study, we used bimolecular fluorescence complementation and in vitro pull-down assays to investigate the supramolecular organization of the CSLC4, XXT1, XXT2, and XXT5 proteins in Arabidopsis protoplasts. Quantification of bimolecular fluorescence complementation fluorescence by flow cytometry allowed us to perform competition assays that demonstrated the high probability of protein-protein complex formation in vivo and revealed differences in the abilities of these proteins to form multiprotein complexes. Results of in vitro pull-down assays using recombinant proteins confirmed that the physical interactions among XXTs occur through their catalytic domains. Additionally, coimmunoprecipitation of XXT2YFP and XXT5HA proteins from Arabidopsis protoplasts indicated that while the formation of the XXT2-XXT2 homocomplex involves disulfide bonds, the formation of the XXT2-XXT5 heterocomplex does not involve covalent interactions. The combined data allow us to propose that the proteins involved in xyloglucan biosynthesis function in a multiprotein complex composed of at least two homocomplexes, CSLC4-CSLC4 and XXT2-XXT2, and three heterocomplexes, XXT2-XXT5, XXT1-XXT2, and XXT5-CSLC4.

Published
2012

3. Xyloglucan Xylosyltransferases XXT1, XXT2, and XXT5 and the Glucan Synthase CSLC4 Form Golgi-Localized Multiprotein Complexes1[W][OA].

Author

Yi Hsiang Chou, Pogorelko, Gennady, and Zabotina, Olga A.

Subjects
*XYLOGLUCANS, *GLUCANS, *GLUCAN synthase, *GOLGI apparatus, *PLANT physiology, *ARABIDOPSIS thaliana, *PHYSIOLOGY
Abstract

Xyloglucan is the major hemicellulosic polysaccharide in the primary cell walls of most vascular dicotyledonous plants and has important structural and physiological functions in plant growth and development. In Arabidopsis (Ambidopsis thaliana), the 1,4-β-glucan synthase, Cellulose Synthase-Like C4 (CSLC4), and three xylosyltransferases, XXT1, XXT2, and XXT5, act in the Golgi to form the xylosylated glucan backbone during xyloglucan biosynthesis. However, the functional organization of these enzymes in the Golgi membrane is currently unknown. In this study, we used bimolecular fluorescence complementation and in vitro pull-down assays to investigate the supramolecular organization of the CSLC4, XXT1, XXT2, and XXT5 proteins in Arabidopsis protoplasts. Quantification of bimolecular fluorescence complementation fluorescence by flow cytometry allowed us to perform competition assays that demonstrated the high probability of protein-protein complex formation in vivo and revealed differences in the abilities of these proteins to form multiprotein complexes. Results of in vitro pull-down assays using recombinant proteins confirmed that the physical interactions among XXTs occur through their catalytic domains. Additionally, coimmunoprecipitation of XXT2YFP and XXT5HA proteins from Arabidopsis protoplasts indicated that while the formation of the XXT2-XXT2 homocomplex involves disulfide bonds, the formation of the XXT2-XXT5 heterocomplex does not involve covalent interactions. The combined data allow us to propose that the proteins involved in xyloglucan biosynthesis function in a multiprotein complex composed of at least two homocomplexes, CSLC4-CSLC4 and XXT2-XXT2, and three heterocomplexes, XXT2-XXT5, XXT1-XXT2, and XXT5-CSLC4. [ABSTRACT FROM AUTHOR]

Published
2012
Full Text
View/download PDF

4. Xyloglucan Xylosyltransferases XXT1, XXT2, and XXT5 and the Glucan Synthase CSLC4 Form Golgi-Localized Multiprotein Complexes1[W][OA].

Author

Yi Hsiang Chou, Pogorelko, Gennady, and Zabotina, Olga A.

Subjects
XYLOGLUCANS, GLUCANS, GLUCAN synthase, GOLGI apparatus, PLANT physiology, ARABIDOPSIS thaliana, PHYSIOLOGY
Abstract

Xyloglucan is the major hemicellulosic polysaccharide in the primary cell walls of most vascular dicotyledonous plants and has important structural and physiological functions in plant growth and development. In Arabidopsis (Ambidopsis thaliana), the 1,4-β-glucan synthase, Cellulose Synthase-Like C4 (CSLC4), and three xylosyltransferases, XXT1, XXT2, and XXT5, act in the Golgi to form the xylosylated glucan backbone during xyloglucan biosynthesis. However, the functional organization of these enzymes in the Golgi membrane is currently unknown. In this study, we used bimolecular fluorescence complementation and in vitro pull-down assays to investigate the supramolecular organization of the CSLC4, XXT1, XXT2, and XXT5 proteins in Arabidopsis protoplasts. Quantification of bimolecular fluorescence complementation fluorescence by flow cytometry allowed us to perform competition assays that demonstrated the high probability of protein-protein complex formation in vivo and revealed differences in the abilities of these proteins to form multiprotein complexes. Results of in vitro pull-down assays using recombinant proteins confirmed that the physical interactions among XXTs occur through their catalytic domains. Additionally, coimmunoprecipitation of XXT2YFP and XXT5HA proteins from Arabidopsis protoplasts indicated that while the formation of the XXT2-XXT2 homocomplex involves disulfide bonds, the formation of the XXT2-XXT5 heterocomplex does not involve covalent interactions. The combined data allow us to propose that the proteins involved in xyloglucan biosynthesis function in a multiprotein complex composed of at least two homocomplexes, CSLC4-CSLC4 and XXT2-XXT2, and three heterocomplexes, XXT2-XXT5, XXT1-XXT2, and XXT5-CSLC4. [ABSTRACT FROM AUTHOR]

Published
2012
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results