caa a22 4500 397555148 CHVBK 20180308164759.0 cr unu---uuuuu 161202e199602 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021250 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021250 Isolation and Partial Characterization of N-Acetyl-D-Galactosamine-Binding Lectins from Epiphragmophora trenquelleonis Snail [Elektronische Daten] [Leonardo Castagna, Jorge Zarzur, Miguel Filipetti, Carlos Landa] 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. © by the Journal of Biochemistry Regular Papers nationallicence albumin gland nationallicence Epiphragmophora trenquelleonis snail nationallicence invertebrate lectins nationallicence N-acetyl-D-galactosamine-binding lectins nationallicence snail nationallicence Castagna Leonardo Centro de Investigations en Qulmica Biológica (CIQUIBIC-CONICET), Departamento de Qulmica Biológica, Facultad de dencias Qutmicas, Uniuersidad Nacional de Córdoba CC #61, Ciudad Universitaria, 5016 Córdoba, Argentina aut Zarzur Jorge Laboratorio de Hemoderivados, Universidad Nacional de Córdoba Córdoba, Argentina aut Filipetti Miguel Laboratorio de Hemoderivados, Universidad Nacional de Córdoba Córdoba, Argentina aut Landa Carlos Centro de Investigations en Qulmica Biológica (CIQUIBIC-CONICET), Departamento de Qulmica Biológica, Facultad de dencias Qutmicas, Uniuersidad Nacional de Córdoba CC #61, Ciudad Universitaria, 5016 Córdoba, Argentina aut The Journal of Biochemistry Oxford University Press 119/2(1996-02), 372-377 0021-924X 119:2<372 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021250 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021250 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Castagna Leonardo Centro de Investigations en Qulmica Biológica (CIQUIBIC-CONICET), Departamento de Qulmica Biológica, Facultad de dencias Qutmicas, Uniuersidad Nacional de Córdoba CC #61, Ciudad Universitaria, 5016 Córdoba, Argentina aut NATIONALLICENCE 700 1- Zarzur Jorge Laboratorio de Hemoderivados, Universidad Nacional de Córdoba Córdoba, Argentina aut NATIONALLICENCE 700 1- Filipetti Miguel Laboratorio de Hemoderivados, Universidad Nacional de Córdoba Córdoba, Argentina aut NATIONALLICENCE 700 1- Landa Carlos Centro de Investigations en Qulmica Biológica (CIQUIBIC-CONICET), Departamento de Qulmica Biológica, Facultad de dencias Qutmicas, Uniuersidad Nacional de Córdoba CC #61, Ciudad Universitaria, 5016 Córdoba, Argentina aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/2(1996-02), 372-377 0021-924X 119:2<372 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford