Isolation and partial characterization of N-acetyl-D-galactosamine-binding lectins from Epiphragmophora trenquelleonis snail. 1996

L Castagna, and J Zarzur, and M Filipetti, and C Landa
Departamento de Química Biológica, Universidad Nacional de Córdoba, Argentina.

A human blood type A hemagglutinating activity was detected in albumin gland extracts of Epiphragmophora trenquelleonis snail separated by GalNAc-agarose affinity chromatography, of which two N-acetyl-D-galactosamine-binding lectins in the extracts were ETL1 was displaced from the affinity column with 1 mM GalNAc, and ETL2 with 20 mM GalNAc. Both lectins agglutinated specifically human blood type A and AB erythrocytes, but not type B and O erythrocytes. Gel filtration chromatography gave a native molecular weight of about 59 kDa for ETL1 and about 54 kDa for ETL2. On SDS-PAGE under nonreducing conditions, ETL1 showed two protein subunits of about 29 and 27 kDa, while ETL2 showed three protein subunits of about 27, 24, and 22 kDa. On SDS-PAGE under reducing conditions, both lectins showed four protein subunits of 17, 16, 12, and 11 kDa. By Western blot analyses developed with biotin-labeled lectins, N-linked oligosaccharides were detected in the 17- and 16-kDa protein subunits of ETL1 and ETL2, and in the 12-kDa protein subunit of ETL2. O-linked oligosaccharides were detected only in the 11-kDa protein subunit of ETL1 and ETL2. On isoelectric focusing both lectins exhibited microheterogeneity: ETL1 focused as three protein bands with pIs in the range of 5.6-6.0, while ETL2 focused as four protein bands with pIs in the range of 6.8-7.4. We suggest that native ETL1 and ETL2 are glycoprotein complexes with molecular weights of 59-54 kDa, composed of two 29-22-kDa nonreduced protein subunits held together by noncovalent hydrophobic interactions. Each of the nonreduced protein subunits seems to be composed of two 17-11-kDa reduced protein subunits held together by interchain disulfide linkages. The main differences between ETL1 and ETL2 could be due to different posttranslational modifications or to the relative contribution of one or more of their protein subunits.

UI MeSH Term Description Entries
D007525 Isoelectric Focusing Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point. Electrofocusing,Focusing, Isoelectric
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D006386 Hemagglutination Tests Sensitive tests to measure certain antigens, antibodies, or viruses, using their ability to agglutinate certain erythrocytes. (From Stedman, 26th ed) Hemagglutination Test,Test, Hemagglutination,Tests, Hemagglutination
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000116 Acetylgalactosamine The N-acetyl derivative of galactosamine. 2-Acetamido-2-D-galactopyranose,2-Acetamido-2-Deoxy-D-Galactose,2-Acetamido-2-Deoxygalactose,N-Acetyl-D-Galactosamine,2 Acetamido 2 D galactopyranose,2 Acetamido 2 Deoxy D Galactose,2 Acetamido 2 Deoxygalactose,N Acetyl D Galactosamine
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012908 Snails Marine, freshwater, or terrestrial mollusks of the class Gastropoda. Most have an enclosing spiral shell, and several genera harbor parasites pathogenic to man. Snail
D037102 Lectins Proteins that share the common characteristic of binding to carbohydrates. Some ANTIBODIES and carbohydrate-metabolizing proteins (ENZYMES) also bind to carbohydrates, however they are not considered lectins. PLANT LECTINS are carbohydrate-binding proteins that have been primarily identified by their hemagglutinating activity (HEMAGGLUTININS). However, a variety of lectins occur in animal species where they serve diverse array of functions through specific carbohydrate recognition. Animal Lectin,Animal Lectins,Isolectins,Lectin,Isolectin,Lectin, Animal,Lectins, Animal

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