Crystals of the Chlamydomonas reinhardtii cell wall: polymerization, depolymerization, and purification of glycoprotein monomers

Two of the three major outer layers of the Chlamydomonas reinhardtii cell wall (W6 and W4) can be solubilized from living cells with sodium perchlorate or other chaotropes and will repolymerize in vitro to form milligram amounts of wall crystals. Conditions for optimal crystalization are presented, and conditions that fail to induce polymerization are exploited to maintain monomers in aqueous solution for ion-exchange chromatography. The four major glycoproteins of the complex (GP1, 1.5, 2, and 3) have in this way been purified to apparent homogeneity and have been characterized morphologically by transmission electron microscopy using the quick-freeze, deep-etch technique and by amino acid composition. Three of the four are hydroxyproline-rich species that copolymerize to form the W6 layer. The fourth (GP1.5) is a glycine-rich species that binds to the interior of the in vitro crystal; it is apparently equivalent to the granules within the W4 layer in situ.

[1]  D. Kirk,et al.  The extracellular matrix of Volvox: a comparative study and proposed system of nomenclature. , 1986, Journal of cell science.

[2]  U. Goodenough,et al.  The Chlamydomonas cell wall and its constituent glycoproteins analyzed by the quick-freeze, deep-etch technique , 1985, The Journal of cell biology.

[3]  S. Imam,et al.  The Chlamydomonas cell wall: characterization of the wall framework , 1985, The Journal of cell biology.

[4]  J. Heuser,et al.  Structure of the Chlamydomonas agglutinin and related flagellar surface proteins in vitro and in situ , 1985, The Journal of cell biology.

[5]  T. Saito,et al.  Cell wall lytic enzyme released by mating gametes of Chlamydomonas reinhardtii is a metalloprotease and digests the sodium perchlorate-insoluble component of cell wall. , 1985, The Journal of biological chemistry.

[6]  K. Jarnagin,et al.  The human insulin receptor cDNA: The structural basis for hormone-activated transmembrane signalling , 1985, Cell.

[7]  K. Roberts,et al.  Cell Wall Glycoproteins: Structure and Function , 1985, Journal of Cell Science.

[8]  U. Goodenough,et al.  Structural comparison of purified dynein proteins with in situ dynein arms. , 1984, Journal of molecular biology.

[9]  G. Ronnett,et al.  Role of glycosylation in the processing of newly translated insulin proreceptor in 3T3-L1 adipocytes. , 1984, Journal of Biological Chemistry.

[10]  R. Mecham,et al.  Chlamydomonas agglutinin is a hydroxyproline-rich glycoprotein. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Heuser,et al.  Procedure for freeze-drying molecules adsorbed to mica flakes. , 1983, Journal of molecular biology.

[12]  B. Monk,et al.  Sexual agglutinins from the Chlamydomonas flagellar membrane. Partial purification and characterization. , 1982, The Journal of biological chemistry.

[13]  R. Mecham,et al.  Antigenicity of elastin: characterization of major antigenic determinants on purified insoluble elastin. , 1982, Biochemistry.

[14]  J. Morrissey,et al.  Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. , 1981, Analytical biochemistry.

[15]  R. Timpl,et al.  Shift in collagen type as an early response to induction of the metanephric mesenchyme , 1981, The Journal of cell biology.

[16]  G. Carpenter,et al.  Epidermal growth factor-receptor-protein kinase interactions. Co-purification of receptor and epidermal growth factor-enhanced phosphorylation activity. , 1980, The Journal of biological chemistry.

[17]  J. Heuser.  Three-dimensional visualization of coated vesicle formation in fibroblasts , 1980, The Journal of cell biology.

[18]  A. Fulton.  Colonial development in Pandorina morum. II. Colony morphogenesis and formation of the extracellular matrix. , 1978, Developmental biology.

[19]  K. Roberts,et al.  The crystalline glycoprotein cell wall of the green alga Chlorogonium elongatum: a structural analysis. , 1976, Journal of cell science.

[20]  K. Roberts,et al.  Self-assembly of a plant cell wall in vitro. , 1975, Journal of molecular biology.

[21]  B. Lindman,et al.  Anion binding properties of human serum albumin from halide ion quadrupole relaxation. , 1975, Biochemistry.

[22]  K. Roberts,et al.  Crystalline glycoprotein cell walls of algae: their stucture, composition and assembly. , 1974, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[23]  K. Roberts,et al.  Structure, composition and morphogenesis of the cell wall of Chlamydomonas reinhardi. I. Ultrastructure and preliminary chemical analysis. , 1972, Journal of ultrastructure research.

[24]  P. V. Hippel,et al.  Ion effects on the solution structure of biological macromolecules , 1969 .

[25]  G. Carpenter,et al.  Epidermal growth factor-receptor-protein kinase interactions. , 1981, Progress in clinical and biological research.

[26]  P. Bornstein,et al.  Structurally distinct collagen types. , 1980, Annual review of biochemistry.

[27]  J. Clamp.  4 – Structure and Function of Glycoproteins , 1975 .

[28]  P. W. Kent.  Structure and function of glycoproteins. , 1967, Essays in biochemistry.