Immunological identification of a putative precursor of the insoluble glycoprotein framework of the Chlamydomonas [Chlorophyceae] cell wall

• Published in: Plant and cell physiology Vol. 37; no. 1; pp. 91 - 101
• Main Authors: Voigt, J. (Technische Univ. Braunschweig (Germany)), Liebich, I, Hinkelmann, B, Kiess, M
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01-01-1996
• Subjects: Amino Acid Sequence; amino acid sequences; Animals; Antibodies; Biological and medical sciences; Blotting, Western; Cell wall; Cell Wall - metabolism; cell wall components; Chlamydomonas reinhardtii; Chlamydomonas reinhardtii - metabolism; CHLOROPHYCEAE; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; GLICOPROTEINAS; GLYCOPROTEINE; Glycoproteins; Glycosylation; IDENTIFICACION; IDENTIFICATION; immunoassays; Insoluble framework; Membrane Glycoproteins - analysis; Membrane Glycoproteins - biosynthesis; Membrane Glycoproteins - chemistry; Metabolism; Molecular Sequence Data; Molecular Weight; Nitrogen metabolism; PARED CELULAR; PAROI CELLULAIRE; Peptide Fragments - chemistry; Plant physiology and development; Precursor; precursors; protein content; Protein Precursors - analysis; Protein Precursors - biosynthesis; Protein Precursors - chemistry; solubility; TECHNIQUE IMMUNOLOGIQUE; TECNICAS INMUNOLOGICAS; Characterization; Antibody; Algae; Chlamydomonas reinhardtii; Chlorophycophyta; Glycoproteins; Biosynthesis precursor; Immunofluorescence; Cell wall; Thallophyta
• ISSN: 0032-0781; 1471-9053
• DOI: 10.1093/oxfordjournals.pcp.a028919

To identify precursors of the insoluble glycoprotein framework of the Chlamydomonas cell wall, a polyclonal antibody was raised against the mixture of polypeptides released from the insoluble wall fraction by chemical deglycosylation. This antibody preferentially cross-reacted with a "150 kDa" salt-soluble cell wall glycoprotein. The conclusion that this "150 kDa" glycoprotein is a putative precursor of the insoluble cell wall fraction was corroborated by the results of pulse-chase experiments and by experiments with antibodies raised against the "150 kDa" salt-soluble glycoprotein and against its 100 kDa deglycosylation product, respectively. Whereas the antibody against the "150 kDa" glycoprotein preferentially recognized carbohydrate side chains, the antibody against its 100 kDa deglycosylation product was found to have essentially the same specificity towards glycosylated and deglycosylated cell wall components as the antibody against the deglycosylation products of the insoluble wall fraction. Furthermore, the antibody against the deglycosylated, insoluble wall fraction recognized almost the same set of peptide fragments derived by V8 protease treatment from the "150 kDa" salt-soluble cell wall glycoprotein and its 100 kDa deglycosylation product, respectively, as the antibody against the 100 kDa deglycosylated cell wall polypeptide