Purification and characterization of Silurus asotus (catfish) roe lectin.
A rhamnose-binding lectin isolated from Silurus asotus (catfish) roe by DEAE-cellulose ion exchange and galactose-Sepharose affinity chromatographies predominantly agglutinated human type B and rabbit erythrocytes. S. asotus lectin (SAL) also agglutinated sarcoma 180 ascites carcinoma cells, but not AH109A cells. The most effective saccharide in hemagglutination inhibition assay was L-rhamnose. The monosaccharides possessing steric similarity to the hydroxyl group orientation at C2 and C4 of the pyranose ring structure of L-rhamnose, such as L-mannose and L-lyxose, were also effective. The molecular weight of SAL was determined to be 38000 by size exclusion chromatography on TSK gel G3000SW and 33000 by SDS-polyacrylamide gel electrophoresis under reducing conditions. SAL did not require a Ca2+ ion or free thiol group for its agglutination activity. The N-terminal 29 amino acid sequence was determined by a gas-phase sequencer as follows, ANMITCYGDVQKLHXETGLIIVKSXLYGR (X: not determined). It has no homology to the sequences of well known vertebrate lectins.
Hosono
M
M
Cancer Research Institute, Tohoku College of Pharmaceutical Sciences, Sendai, Japan.
Kawauchi
H
H
Nitta
K
K
Takayanagi
Y
Y
Shiokawa
H
H
Mineki
R
R
Murayama
K
K
D000371:Agglutination; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002397:Catfishes; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006384:Hemagglutination; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D008358:Mannose / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D012210:Rhamnose / Q000378:metabolism*; D016415:Sequence Alignment; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D037102:Lectins; D008358:Mannose; D012210:Rhamnose