Purification and characterization of the agglutinins from the sponge Axinella polypoides and a study of their combining sites

Bretting, Hagen; Kabat, Elvin A.

doi:10.1021/bi00660a011

Cited by 99 publications

(43 citation statements)

References 45 publications

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“…The native molecular weight of both lectins was 63,000, thus suggesting that both lectins in the native state exhibit a homotetrameric structure. That is a common structural feature from some of the sponge lectins characterized to date, along with the acidic pI (4,5,(21)(22)(23)(24)(25). The tetrameric structure was further confirmed by the Scatchard analysis, which indicated the presence of four homogeneous carbohydrate binding sites per native molecule of lectin.…”

Section: Discussionmentioning

confidence: 97%

Lectins from Tropical Sponges

Miarons

Fresno

2000

Journal of Biological Chemistry

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Only a few animal phyla have been screened for the presence and distribution of lectins. Probably the most intensively studied group is the mollusk. In this investigation, 22 species from 12 families of tropical sponges collected in Los Roques National Park (Venezuela) were screened for the presence of lectins. Nine saline extracts exhibited strong hemagglutinating activity against pronase-treated hamster red blood cells; five of these reacted against rabbit red blood cells, four with trypsintreated bovine red blood cells, and five with human red blood cells regardless of the blood group type. Extracts from the three species studied from genus Aplysina (archeri, lawnosa, and cauliformis) were highly reactive and panagglutinating against the panel of red blood cells tested. The lectins from A. archeri and A. lawnosa were purified to homogeneity by ammonium sulfate fractionation, affinity chromatography on p-aminobenzyl-␤-1-thiogalactopyranoside-agarose, and gel filtration chromatography. Both lectins exhibited a native molecular mass of 63 kDa and by SDS-polyacrylamide gel electrophoresis under reducing conditions have an apparent molecular mass of 16 kDa, thus suggesting they occur as homotetramers. The purified lectins contain 3-4 mol of divalent cation per molecule, which are essential for their biological activity. Hapten inhibition of hemagglutination was carried out to define the sugar binding specificity of the purified A. archeri lectin. The results indicate a preference of the lectin for nonreducing ␤-linked D-Gal residues being the best inhibitors of red blood cells binding methyl-␤-D-Gal and thiodigalactoside (Gal␤1-4-thiogalactopyranoside). The behavior of several glycans on immobilized lectin affinity chromatography confirmed and extended the specificity data obtained by hapten inhibition.In animals, only a few phyla have been screened for the presence and distribution of lectins. In particular, the number of lectins that have isolated from invertebrate organisms is quite small as compared with the great variety of lectins isolated from plant origin and have been limited to those partially characterized from mollusks and crustaceans. Since the discovery of hemagglutinins in sponges by Dodd et al. (1), there have been some reports on the partial characterization (serological and immunoelectrophoretic properties) of lectins from the oldest multicellular animals (2, 3) from the Mediterranean Sea or Japan (4 -8). However, their properties and specificities have not been clearly defined. In this report, we present data on an investigation undertaken to search for novel lectins in marine organisms, which could show unique properties. In addition, we describe the purification and characterization of the lectins present in two species of tropical sponges, Aplysina lawnosa and Aplysina archeri. We present information on the nature and specificity of their combining site as examined by hapten inhibition experiments and affinity chromatography.

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Section: Discussionmentioning

confidence: 97%

Lectins from Tropical Sponges

Miarons

Fresno

2000

Journal of Biological Chemistry

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“…D-Galactose is a potent inhibitor of both Axinella and Cerianthus agglutinins and these agglutinins also demonstrate some degree of {3-anomeric specificity (Baldo et al 1977a(Baldo et al , 1977b. The Axinella agglutinins are best inhibited by terminal non-reducing D-galactose glycosidically linked {3-(1'-+6) (Bretting and Kabat 1976;Eichmann et al 1976).…”

Section: Discussionmentioning

confidence: 99%

Reaction of Some Invertebrate and Plant Agglutinins and a Mouse Myeloma Anti-Galactan Protein With an Arabinogalactan From Wheat

Baldo¹,

Neukom²,

Stone³

et al. 1978

Aust. Jnl. Of Bio. Sci.

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Plant, invertebrate and vertebrate proteins which show anti-galactan combining specificities were used in precipitation and inhibition studies with arabinogalactan preparations from wheat and ryegrass (Lolium multiflorum). Of the agglutinins studied, only mouse anti-galactan myeloma protein J539 showed strong reactivity with wheat arabinoglactan-peptide. Weak reactions were observed with the agglutinins from the clam Tridacna maxima, the sponge Axinella polypoides and the anemone Cerianthus membranaceus. No reactions were detected with lectins from the plants Abrus precatorius and Ricinus communis. Reactions readily occurred between Lolium arabinogalactan-protein and the invertebrate and vertebrate agglutinins. Removal of terminal arabinosyl residues from the wheat and Lotium arabinogalactans either by mild acid hydrolysis or by treatment with an arabinofuranosidase increased the reactivity of both peptidoglycans with all of the agglutinins examined except the Ricinus RCA l lectin. Results obtained with wheat arabinogalactan indicate that few D-galactose units are terminal and available for reaction. The difference in reactivities between the wheat and Lotium arabinogalactans may be due to the differences in the galactose:arabinose ratios or to differences in linkage of the galactosyl residues on the two peptidoglycans, or both. Results indicate that the mouse anti-galactan could be a useful reagent for the subcellular localization of wheat arabinogalactan and that tridacnin and Axinella agglutinins could be used to localize the arabinogalactan in L. multiflorum cells.

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“…It is a mixture of two agglutinins which are inhibited by glyco-substances with terminal/)-galactosyl residues, preferably 0-1 -+ 6 linked (11,12). '…”

Section: Lectin From Abrus Precatoriusmentioning

confidence: 99%

Additional Precipitation Reactions of Lectins with Human Serum Glycoproteins

Uhlenbruck¹,

Baldo²,

Steinhausen³

et al. 1978

Clinical Chemistry and Laboratory Medicine

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Summary: Highly purified human serum glycoproteins were treated with neuraminidase and examined for their cross reaction with several lectins with anti-galactosyl specificity: )3-.D-galactosyl structures are thought to be the main terminal sugar residues that become attached de novo after removal of neuraminic acid. The following lectins were tested: Tridacnin from the bivalve clams Tridacna maxima and Tridacna gigas, the agglutinin from the sponge Axinella polypoides 9 the lectin from the roach Rutilus rutilus and the plant lectins from Ricinus communis, Ononis spinosa, Glycine soja zndAbrus precatorius. In agar gel diffusion, these purified and precipitating lectins gave more or less strong or negative results against the different neuraminidase-treated serum glycoproteins, thus indicating subtle differences with respect to their anti-galactosyl combining specificity. On the other hand, serum glycoproteins which reacted with the same lectin, did not always show complete identity lines. Finally, as revealed by these lectins, the carbohydrate moiety of serum glycoproteins may reflect a complex and broad spectrum of heterogeneity. This could lead to a more detailed understanding of the topographical and steric arrangement of the chemical structure and of the biological role of carbohydrate groups in these glycosubstances. Zusätzliche Fällungsreaktionen von Lectinen mit Glykoproteinen aus MenschenserenZusammenfassung: Menschliche Seruin-Glykoproteine wurden mit Neuraminidase behandelt und im Hinblick auf ihre Kreuzreaktionen mit verschiedenen Lectinen mit Anti-Galactosyl-Spezifität untersucht. Es wird angenommen, daß )3--D-Galactosyl-Strukturen in der Regel diejenigen Zuckerreste sind, welche nach Entfernung der Neuraminsäure endständig werden. Folgende Lectine wurden getestet: Tridacnin aus den Muscheln Tridacna maxima und Tridacna gigas, das Agglutinin vom Schwamm Axinella polypoides, das Lectin aus der Plötze Rutilus rutilus und die Pflanzenlectine oasRicinus communis, Ononis spinosa 9 Glycine sofa undAbrus precatorius. Die gereinigten, präzipitierenden Lectine gaben mehr oder weniger starke Präzipitate, wenn sie im Agargel gegen verschiedene, Neuraminidase-behandelte SerumGlykoproteine diffundierten. Auf diese Weise konnten feine Unterschiede bezüglich der Anti-Galactosyl-Spezifität aufgezeigt werden, während andererseits Serum^Glykoproteine, die mit demselben Lectin reagierten, nicht immer vollständige Identitätslinien ergäben. Die Ergebnisse deuten an, wie komplex und heterogen der Kohlenhydratanteil dieser Serum-Glykoproteine ist, wenn man um mit mehreren Lectinen untersucht. Aber nur so gewinnt man ein detailliertes Bild von der topographischen und sterischen Anordnung dieser Kohlenhydratketten in diesen Glykosubstanzeü und kann etwas über ihre Chemie und biologische Bedeutung in Erfahrung bringen.

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Purification and characterization of the agglutinins from the sponge Axinella polypoides and a study of their combining sites

Cited by 99 publications

References 45 publications

Lectins from Tropical Sponges

Lectins from Tropical Sponges

Reaction of Some Invertebrate and Plant Agglutinins and a Mouse Myeloma Anti-Galactan Protein With an Arabinogalactan From Wheat

Additional Precipitation Reactions of Lectins with Human Serum Glycoproteins

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