Heterogeneity of lobster agglutinins. I. Purification and physicochemical characterization

Hall, James L.; Rowlands, David T.

doi:10.1021/bi00701a028

Cited by 105 publications

(28 citation statements)

References 23 publications

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“…These peptides, released by papain digestion of hog gastric mucin and purified by ethanol and cetylpyridinium chloride precipitation, appear to contain the full complement of sugars found on glycoproteins (27). The lectins ConA, WGA, soybean agglutinin, RCAs, Lotus lectin, and Ulex lectin, previously mentioned in this paper, as well as lima bean agglutinin, Limulin (26), and a lectin from lobster hemolymph (10,11), have been found by us to bind to these insolubilized glycopeptides and can be eluted by their hapten sugars or by increases in ionic strength, as in the case of Limulin and lobster lectin (10,11,26). Other agglutinins have also been reported to bind to hog gastric mucin.…”

Section: Discussionsupporting

confidence: 51%

Distribution of cell surface saccharides on pancreatic cells

Mf¹,

Jd²

1979

The Journal of Cell Biology

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We describe here a simple, general procedure for the purification of a variety of lectins, and for the preparation of lectin4erritin conjugates of defined molar composition and binding properties to be used as probes for cell surface saccharides. The technique uses a "universal" affinity column for lectins and their conjugates, which consists of hog sulfated gastric mucin glycopeptides covalently coupled to agarose. The procedure involves: (a) purification of lectins by chromatography of aqueous extracts of seeds or other lectin-containing fluids over the affinity column, followed by desorption of the desired lectin with its hapten sugar; (b) iodination of the lectin to serve as a marker during subsequent steps; (c) conjugation of lectin to ferritin with glutaraldehyde; (d) collection of active lectin-ferritin conjugates by affinity chromatography; and (e) separation of monomeric lectin-ferritin conjugates from larger aggregates and unconjugated lectin by gel chromatography. Based on radioactivity and absorbancy at 310 nm for lectin and ferritin, respectively, the conjugates consist of one to two molecules of lectin per ferritin molecule. Binding studies of native lectins and their ferritin conjugates to dispersed pancreatic acinar cells showed that the conjugation procedure does not significantly alter either the affinity constant of the lectin for its receptor on the cell surface or the number of sites detected.KEY WORDS lectin affinity purificationmucin peptides 9 lectin-ferritin conjugates conjugate binding characteristics Because of their high specificity of binding to the carbohydrate moieties of glycoproteins and glycolipids, lectins have proven to be powerful tools for examination of the architecture of the plasmalemma, comparison of the surface saccbaride patterns of normal and transformed cells, and detection of cell surface changes during development (15). For these purposes, a variety of lectins with different sugar specificities have been purified by either conventional procedures or by affinity chromatographic (15) techniques applicable to individual iectins. In addition, several general affinity adsorbents have been proposed for isolation of groups of lectins. For example, fixed erythrocytes J. CELL BIOLOGY 9 The Rockefeller University Press -0021-

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Section: Discussionsupporting

confidence: 51%

Distribution of cell surface saccharides on pancreatic cells

Mf¹,

Jd²

1979

The Journal of Cell Biology

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“…Recent work on the lobster's mechanisms for resistance against disease has concent-rated on elucidating the agglutinin and opsonin subject areas by using materials separated chromatographically from lobster hemolyrnph (Hall & Rowlands, 1974;Hartman et al, 1978;Vanderwall et al, 1981;Goldenberg & Greenberg, 1983). Goldenberg & Greenberg were able to demonstrate differences in the properties of the agglutinin versus the opsonin, but were not prepared to state definitively that these were completely separate molecular entities.…”

Section: Epibiontic Growthmentioning

confidence: 99%

Lobster diseases

Stewart

1984

Helgolander Meeresunters

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“…5 ,31) Structural analysis of BRAs will help us to analyze the structural basis for their different affinities against sugars and understand the origin and function of molecular diversity.…”

Section: Discussionmentioning

confidence: 99%