title: Molecular cloning of cDNAs encoding the protein backbones of arabinogalactan-proteins from the filtrate of suspension-cultured cells of Pyrus communis and Nicotiana alata.
abstract: This paper reports the isolation of cDNAs encoding the protein backbone of two arabinogalactan-proteins (AGPs), one from pear cell suspension cultures (AGPPc2) and the other from suspension cultures of Nicotiana alata (AGPNa2). The proteins encoded by these cDNAs are quite different from the 'classical' AGP backbones described previously for AGPs isolated from pear suspension cultures and extracts of N. alata styles. The cDNA for AGPPc2 encodes a 294 amino acid protein, of which a relatively short stretch (35 amino acids) is Hyp/Pro rich; this stretch is flanked by sequences which are dominated by Asn residues. Asn residues are not a feature of the 'classical' AGP backbones in which Hyp/Pro, Ser, Ala and Thr account for most of the amino acids. The cDNA for AGPNa2 encodes a 437 amino acid protein, which contains two distinct domains: one rich in Hyp/Pro, Ser, Ala, Thr and the other rich in Asn, Tyr and Ser. The composition and sequence of the Pro-rich domain resembles that of the 'classical' AGP backbone. The Asn-rich domains of the two cDNAs described have no sequence similarity; in both cases they are predicted to be processed to give a mature backbone with a composition similar to that of the 'classical' AGPs. The study shows that different AGPs can differ in the amino acid sequence in the protein backbone, as well as the composition and sequence of the arabinogalactan side-chains. It also shows that differential expression of genes encoding AGP protein backbones, as well as differential glycosylation, can contribute to the tissue specificity of AGPs.
authors.affiliation: CRC for Industrial Plant Biopolymers and Plant Cell Biology Research Centre, School of Botany, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: S L
authors.initials: SL
authors.lastname: Mau
authors.forename: C G
authors.initials: CG
authors.lastname: Chen
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Pu
authors.forename: R L
authors.initials: RL
authors.lastname: Moritz
authors.forename: R J
authors.initials: RJ
authors.lastname: Simpson
authors.forename: A
authors.initials: A
authors.lastname: Bacic
authors.forename: A E
authors.initials: AE
authors.lastname: Clarke
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005685:Galactans; D008969:Molecular Sequence Data; D009088:Mucoproteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D059828:Plant Cells; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D010944:Plants / Q000235:genetics*; D010947:Plants, Toxic; D011499:Protein Processing, Post-Translational; D011509:Proteoglycans / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D017421:Sequence Analysis; D014026:Nicotiana / Q000166:cytology / Q000235:genetics; D014197:Trees / Q000166:cytology / Q000235:genetics
chemical_list: D000596:Amino Acids; D018076:DNA, Complementary; D005685:Galactans; D009088:Mucoproteins; D010940:Plant Proteins; D011509:Proteoglycans; C006619:arabinogalactan proteins; C005653:arabinogalactan