Purification and characterization of a novel lectin from a freshwater cyanobacterium, Oscillatoria agardhii.
In the survey of 14 species of laboratory-cultured cyanobacteria for hemagglutinins, we newly detected the activity in two species, Oscillatoria agardhii, strain NIES-204, and Phormidium foveolarum, strain NIES-503. From the extract of O. agardhii, which showed the highest activity with trypsin-treated erythrocytes of rabbit, a lectin was purified to homogeneity by the combination of precipitation with (NH4)2SO4, gel filtration, hydrophobic chromatography and reverse phase chromatography. The purified lectin, designated OAA, was a monomeric protein with an apparent molecular weight of 13,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 16,000 on gel filtration. The amino acid composition was rich in glycine and acidic amino acids. The hemagglutination activity was inhibited by glycoproteins such as yeast mannan, but not by any of the monosaccharides tested. The activity was stable over a wide range of pH (4-11) and at a high temperature of 80 degrees C, and independent on the presence of divalent cations. The features of OAA resembled those of many of lectins from marine macroalgae. The sequence of amino-terminal residues of OAA was determined as ALYNVENQWGGSSAPWNEGG, which was highly homologous to those of lectins from macroalgae of the genus Eucheuma and that of a myxobacterium Myxococcus xanthus hemagglutinin.
Sato
Y
Y
Faculty of Applied Biological Science, Hiroshima University, Japan.
Murakami
M
M
Miyazawa
K
K
Hori
K
K
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000737:chemistry; D001426:Bacterial Proteins; D050260:Carbohydrate Metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002457:Cell Extracts; D002850:Chromatography, Gel; D000458:Cyanobacteria / Q000737:chemistry* / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006023:Glycoproteins / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests; D006388:Hemagglutinins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011817:Rabbits; D000461:Rhodophyta / Q000737:chemistry
D001426:Bacterial Proteins; D002352:Carrier Proteins; D002457:Cell Extracts; D006023:Glycoproteins; D006388:Hemagglutinins; D037102:Lectins; C415125:OAA protein, Oscillatoria agardhii