Cloning and characterization of AtRGP1: a reversibly autoglycosylated Arabidopsis protein implicated in cell wall biosynthesis

• Published in: Plant physiology (Bethesda) 1998-04, Vol.116 (4), p.1339-1349
• Main Authors: Delgado, I.J. (Michigan State University, East Lansing, MI.), Wand, Z, De Rocher, A, Keegstra, K, Raikhel, N.V
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; AMINO ACID SEQUENCES; Antibodies; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins; ARABIDOPSIS THALIANA; BINDING SITES; Biological and medical sciences; BIOSYNTHESIS; Biotechnology; Cell Biology and Signal Transduction; CELL CULTURE; CELL MEMBRANES; CELL SUSPENSIONS; Cell Wall - metabolism; CELL WALLS; CHEMICAL COMPOSITION; CHEMICAL REACTIONS; CLONING; Cloning, Molecular; COMPLEMENTARY DNA; Corn; DNA; DNA, Complementary; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; GALACTOSE; GENBANK/AF005279; GENBANK/AF013627; GENBANK/U31565; GENBANK/U89897; GENE EXPRESSION; GENES; Genes. Genome; Genetic engineering; GENETIC REGULATION; Genetic technics; GENETICS; GLUCOSE; Glycoproteins - genetics; Glycoproteins - metabolism; Glycosylation; HEMICELLULOSE; LOCALIZATION; Membrane Proteins - genetics; Membrane Proteins - metabolism; Methods. Procedures. Technologies; Molecular and cellular biology; Molecular cloning; Molecular genetics; MOLECULAR SEQUENCE DATA; NUCLEOTIDES; Peas; PISUM SATIVUM; Plant Proteins - genetics; Plant Proteins - metabolism; Plants; POLYSACCHARIDES; Polysaccharides - biosynthesis; Proteins; RNA; RNA, Messenger - genetics; RNA, Messenger - metabolism; ROOTS; Sequence Homology, Amino Acid; Solubility; Space life sciences; SUBSTRATES; UDP
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.116.4.1339

A reversibly glycosylated polypeptide from pea (Pisum sativuum) is thought to have a role in the biosynthesis of hemicellulosic polysaccharides. We have investigated this hypothesis by isolating a cDNA clone encoding a homolog of Arabidopsis thaliana, Reversibly Glycosylated Polypeptide-1 (AtRGP1), and preparing antibodies against the protein encoded by this gene. Polyclonal antibodies detect homologs in both dicot and monocot species. The patterns of expression and intracellular localization of the protein were examined. AtRGP1 protein and RNA concentration are highest in roots and suspension-cultured cells. Localization of the protein shows it to be mostly soluble but also peripherally associated with membranes. We confirmed that AtRGP1 produced in Escherichia coli could be reversibly glycosylated using UDP-glucose and UDP-galactose as substrates. Possible sites for UDP-sugar binding and glycosylation are discussed. Our results are consistent with a role for this reversibly glycosylated polypeptide in cell wall biosynthesis, although its precise role is still unknown