A Preliminary Analysis of the Proteins of the Primary Walls of some Plant Cells

King, N. J.; Bayley, S. T.

doi:10.1093/jxb/16.2.294

Cited by 41 publications

(17 citation statements)

References 8 publications

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“…The amino acid and carbohydrate composition of the glycoproteins, however, also showed certain similarities. For example, as noticed before on less purified samples [4,10,25] and supported here, hydroxy amino acids were rather prominent in these macromolecules. I n addition, the rather low contents of acidic amino acids for plant proteins were also noteworthy.…”

Section: Discussionsupporting

confidence: 88%

Fractionation and Characterization of Glycoproteins Containing Hydroxyproline from the Leaves of Vicia faba

Pusztai¹,

Watt²

1969

Eur J Biochem

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A survey was conducted of the distribution, solubility, chemical and some physical properties of the constituents of the leaves of Vicia faba containing hydroxyproline. With aqueous trichloroacetic acid about 30OlO of the total leaf hydroxyproline could be brought into solution. The rest remained insoluble in aqueous or non-aqueous solvents tested under non-hydrolytic conditions. The material soluble in trichloroacetic acid was resolved by phenol-aqueous (pH 8) partitioning into (a) predominantly polysaccharide-type materials with small amounts of firmly bound proteins containing hydroxyproline; (b) into some insoluble material and (c) into glycoproteins and glycolipids containing hydroxyproline soluble in the phenol-rich phase, These last were separated into a t least four cationic, one anionic and one stationary components on continuous high voltage electrophoresis in phenol-acetic acid-water. The four cationic glycoprotein fractions were resolved further on the basis of their size on a Bio Gel P 100 column operated in phenol-acetic acid-water. The stationary component was composed of a mixture of glycolipids and contained strongly bound amino acids including hydroxyproline. Analytical results for the major components obtained in this work clearly indicated the presence in the leaves of Vicia faba of a number of glycoproteins, polysaccharide-protein complexes and glycolipids containing hydroxyproline each with characteristic and different composition, molecular size and solubility. Previous work on cell-wall protein (extensin) was discussed in the light of these results.Since the discovery by Steward and Thompson [l] of hydroxyproline in hydrolysates of tissues of plant origin it is now firmly established that this hydroxy imino acid is widely distributed in the plant kingdom. Its occurrence, exact chemical structure and association with other constituents and structural elements of the plant cell have been extensively reviewed [2-41. Hydroxyproline has been reported to be associated with materials in the pericarps and protective seed coats of plants [5-71 and, more specifically, with structural elements of the walls of cultured plant cells [8,9] and of normally grown plant tissues [lo]. Hydroxyproline of plant tissues has been shown to originate from proline which after incorporation into components of the cell is hydroxylated [ll-131. Free hydroxyproline inhibits both the auxin-induced cell elongation and the formation of protein-bound hydroxyproline [14-171.It will incorporate into plant cells only under special conditions and in small amounts [14,18,19]. I n view of this close association Lamport [3] proposed that hydroxyproline is confined to the cell wall and, consequently, its amount in the plant cell could serve as a measure of the cell wall. Recent studies, in addition to some earlier evidence [4], have however cast doubt on the validity of this view. For example, it has since been claimed that collagen is present in plant nuclei [20]. Similarly, in Avena coleoptiles a t least 20°/0 of the total hydro...

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Section: Discussionsupporting

confidence: 88%

Fractionation and Characterization of Glycoproteins Containing Hydroxyproline from the Leaves of Vicia faba

Pusztai¹,

Watt²

1969

Eur J Biochem

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“…1-owever, the possibility of cytoplasmic contamination of the walls is tunlikely. For example, the absence of particul,ate contamination of cell walls prepared by the procedtire employed has been previouis'ly shown, Utsinlg both the light microscope (3), an'd electron microscope (6). Similarly, the efificacy of the glycerol washing procedture has been stuidied by deliberate contamination of the walls with a solulble enzyme and subsequent repturification (4 protein and of various synthetic and hydrolytic enzym.es in the primary cell wallI (7), and significantly, the unique presence of hydroxyprolline-rich protein,s in the wall (8).…”

Section: Discussionmentioning

confidence: 99%

In Vitro Autolysis of Plant Cell Walls

1967

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“…The cell wall preparations obtained appeared free of particulate contamination in the light microscope (13) and similarly prepared walls from other tissues seemed homogeneous when examined in the electron microscope (12). Reisolation of the walls, after deliberate contamination with a cytoplasmic enzyme, indicated that the preparative procedure used was adequate to remove the added cytoplasmic contamination (10,14).…”

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confidence: 92%

A Partial Characterization of an Autolytically Solubilized Cell Wall Glucan

1971

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(15)(16)(17). In this paper evidence is presented to indicate that wall-localized enzymes solubilize only one of the polysaccharides of the wall. This polysaccharide resembles, in terms of its hydrolysis products, the glucans, lichenan (32) and oat and barley glucan (31). MATERIALS AND METHODSCell Wall Preparation and Incubation. Cell walls were prepared from 5-day corn coleoptile tissue by homogenization in glycerol and filtration through a glass bead filter, as previously described (13). Glycerol was removed, prior to incubation, by washing the walls with 95% ethyl alcohol, then acetone, and then ether, all at -10 to -20 C (17). Solvent-washed cell wall powders were stored at -20 C over anhydrous calcium sulfate (17) and were stable with regard to autolytic capability for at least 1 year. Incubation was at 37 C in water with a wall concentration of 10 mg/ml and with occasional resuspension by shaking. "Zero" time controls were obtained by heating the suspended walls in a boiling water bath immediately after suspension. All incubations were terminated at the indicated time in the same way as the zero time controls. Residual wall was collected by filtration through Millipore filters (0.45 /) or through washed and tared discs of Whatman No. 540. The collected walls were washed with water (3 X 1 ml) and the wash waters were combined and filtrate was frozen and thawed three times to promote retrogradation of solubilized wall components and these then were collected by filtration and weighed. Unless otherwise specified, reagents were of analytical quality and were used without prior purification. Glycerol was distilled in vacuo prior to use. Cellobiose' and gentiobiose were obtained from Nutritional Biochemicals. A sample of laminaribiose was obtained from Professor W. Z. Hassid. Later, samples of laminaribiose were prepared by partial acid hydrolysis of laminaran (Pfaltz and Bauer, Inc.) and the di-and trisaccharides were isolated by chromatography on Sephadex G-10 or G-15. The trisaccharide

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A Preliminary Analysis of the Proteins of the Primary Walls of some Plant Cells

Cited by 41 publications

References 8 publications

Fractionation and Characterization of Glycoproteins Containing Hydroxyproline from the Leaves of Vicia faba

Fractionation and Characterization of Glycoproteins Containing Hydroxyproline from the Leaves of Vicia faba

In Vitro Autolysis of Plant Cell Walls

A Partial Characterization of an Autolytically Solubilized Cell Wall Glucan

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