Isolation of an arabinogalactan protein by lectin affinity chromatography on tridacnin-Sepharose 4B

doi:10.1016/0003-2697(79)90622-5

Analytical Biochemistry

Volume 92, Issue 1, 1 January 1979, Pages 41-45

https://doi.org/10.1016/0003-2697(79)90622-5 Get rights and content

Abstract

An arabinogalactan protein from the style canal of Gladiolus has been isolated by a one-step procedure involving lectin affinity chromatography, using the galactose-binding lectin from the small giant clam, Tridacna maxima, coupled to Sepharose 4B. The lectin binding is calcium-ion dependent, so that the arabinogalactan protein which is bound to the column in the presence of calcium can be eluted by washing the column with a calcium-free buffer. This method has a general utility for the purification of polysaccharides and glycoproteins containing β-linked galactosyl residues.

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Cited by (8)

The Yariv reagent: Behaviour in different solvents and interaction with a gum arabic arabinogalactanprotein
2014, Carbohydrate Polymers
Citation Excerpt :
The role of terminal Araf residues in the interaction is seemingly complex. Both Gleeson, Jermyn, and Clarke (1979) and Akiyama and Kato (1981) showed that the progressive removal of terminal Araf residues by acid hydrolysis led to a loss in complexing ability. Treatment of rose (Rosa glauca) AGP with 12.5 M oxalic acid at 100 °C for 5 h led to a loss in 3,6-linked Gal with a parallel increase in 6-linked Gal and resulted in a product with a greatly diminished interaction with the β-d-Glc Yariv reagent (Komalavilas, Zhu, & Nothnagel, 1991).
The β-d-Glc Yariv reagent is frequently used to isolate and to study the structure of arabinogalactan-proteins with the arabinogalactan type II structure. The present paper describes the aggregation features of the Yariv reagent in water, salt solutions and in organic solvents as determined by NMR, absorption spectroscopy and light scattering experiments. The results indicate that in water the Yariv reagent forms aggregates of up to 300 units and in 1% aqueous NaCl the degree of aggregation is approx. 150. The aggregates are formed both by H-bonds and hydrophobic interactions, the former appearing to be of most importance in water.
The interaction between the Yariv reagent and an AGP fraction from gum arabic, showed a degree of aggregation of the Yariv reagent when using 1% NaCl to be of approx. 150 units, whereas disruption of the aggregate took place in 10% NaCl with an aggregation number of approx. 100.
Partial acid hydrolysis of an AGP from gum Arabic (Acacia Senegal) and analyses of the linkage types remaining indicated that a certain length of (1 → 3)-β-linked galactose units was necessary for binding between the Yariv reagent and the AGP. This is in accordance to what also was recently observed by Kitazawa et al. (2013).
Quantification of arabinogalactan-protein in plant extracts by single radial gel diffusion
1985, Analytical Biochemistry
The amount of arabinogalactan-protein in whole plant extracts can be quantified by single radial diffusion in agarose gels containing a dye known as the β-glucosyl-Yariv reagent which specifically interacts with and precipitates arabinogalactan-proteins. The lower limit of quantification is 0.04 μg of arabinogalactan-protein; gum arabic is used as a standard reference arabinogalactan-protein. In principle, this method can be adapted to measure levels of any dyeprecipitating macromolecule; for example, acidic polysaccharides can be estimated by their binding to the cationic dye Alcian blue.
Carbohydrates
1983, Journal of Chromatography Library
This chapter introduces the use of the newer techniques in the isolation and analysis of carbohydrates and their derivatives, but, within this context, some of the longer-established methods—such as ion-exchange and gel-permeation chromatography—remain relevant, particularly in view of their compatibility with the various automated detection systems now available for chromatographic analysis of carbohydrates. The term column chromatography is applied to many different types of system, in which the separation mechanism may depend upon adsorption, partition, ion exchange, or molecular exclusion, according to the nature of the stationary phase. However, it is becoming increasingly apparent that the boundaries between these various mechanisms are vague, as more than one mechanism is frequently involved in a particular separation, especially in the case of some of the packings now used in high performance liquid chromatography (HPLC). The once important techniques of adsorption chromatography on charcoal–Celite columns and partition chromatography on powdered cellulose are now obsolete as analytical methods, but retain some value for preparative purposes.
Affinity chromatography of arabinogalactan-proteins
1983, Carbohydrate Polymers
Arabino-(1→3), (1→6)-β-d-galactan-proteins (AGPs) and related compounds from Lolium multiflorum (ryegrass) endosperm cell suspension culture, wheat endosperm, larchwood and Gladiolus stigma extract were shown to bind selectively at neutral pH to a column of Sepharose to which the anti-galactan myeloma protein J539 had been covalently linked, Elution was achieved with buffer at pH 3 or with a pulse of p-nitrophenyl-β-d-galactopyranoside at neutral pH. These observations formed the basis for an affinity chromatographic purification of AGPs from natural sources. Some heterogeneity in a ryegrass AGP preparation was indicated by its incomplete elution by d-galactose.
Arabinogalactans in a purified allergen preparation from pollen of timothy (Phleum pratense)
1982, Phytochemistry
The purified allergen preparation representing a certain fraction of an aqueous timothy pollen extractcontained ca. 20% carbohydrate, mainly as arabinose (7%) and galactose (13%). The protein content was 63%. Fractionation on DEAE-Sephadex and Sephadex G-100 gave one neutral and two acidic fractions, all containing protein, arabinose and galactose. The structure of the carbohydrate moiety was investigated by methylation analysis, periodate oxidation and enzyme incubation. The acidic fraction contained (1→6)-linked galactose residues, some being substituted on O-3 with arabinose. The neutral fraction consisted of a more extensively branched arabinogalactan with longer side chains of (1→3)- and (1→5)-linked arabinose. The arabinose was present mainly as α-l-arabinofuranosyl residues. Alkaline degradation and subsequent fractionation indicated the presence of a covalent linkage between hydroxyproline and arabinose. Periodate oxidation or incubation with α-l-arabinofuranosidase did not affect the allergenic activity of the extract.
Arabinogalactans of sexual and somatic tissues of gladiolus and lilium
1980, Phytochemistry
Arabinogalactans are present in extracts of the sexual and somatic tissues of two monocotyledons, Gladiolus and Lilium. The arabinogalactans precipitated from plant extracts by the β-glucosyl artificial antigen (Yariv antigen) show differences in the proportions of the major monosaccharides, galactose and arabinose. Gladiolus stigma and style preparations contain a single arabinogalactan component, but the other tissues examined contained at least two distinct groups of arabinogalactans. These groups can be separated electrophoretically or by lectin affinity chromatography.

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Isolation of an arabinogalactan protein by lectin affinity chromatography on tridacnin-Sepharose 4B

Abstract

Carbohyd. Res

Immunochemistry

Carbohyd. Res

Comp. Biochem. Physiol

Aust. J. Plant Physiol

Phytochemistry

Quart. Rev. Biol

Aust. J. Plant Physiol

Biochem. J