Purification and carbohydrate-binding specificity of Agrocybe cylindracea lectin.
A lectin was isolated from fruiting bodies of Agrocybe cylindracea by two ion-exchange chromatographies and gel filtration on Toyopearl HW55F. The lectin was homogeneous on polyacrylamide gel electrophoresis and its molecular mass was determined to be 30000 by gel filtration, and 15000 by sodium dodecylsulfate polyacrylamide gel electrophoresis, signifying a dimeric protein. Its carbohydrate-binding specificity was investigated both by sugar-hapten inhibition of hemagglutination and by enzyme-linked immunosorbent assay. The inhibition tests showed the affinity of the lectin to be weakly directed toward sialic acid and lactose, and the enhanced affinity toward trisaccharides containing the NeuAc alpha2,3Gal beta-structure. Importantly, the lectin strongly interacted with glycoconjugates containing NeuAc alpha2,3Gal beta1,3GlcNAc-/GalNAc sequences.
Yagi
F
F
Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, Japan.
Miyamoto
M
M
Abe
T
T
Minami
Y
Y
Tadera
K
K
Goldstein
I J
IJ
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001487:Basidiomycota / Q000737:chemistry*; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000494:pharmacology; D002364:Caseins / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005732:Gangliosides / Q000378:metabolism / Q000494:pharmacology; D006001:Glycoconjugates / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006384:Hemagglutination / Q000187:drug effects; D007797:Laminin / Q000378:metabolism; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009844:Oligosaccharides / Q000494:pharmacology; D010940:Plant Proteins / Q000302:isolation & purification / Q000378:metabolism; D017421:Sequence Analysis; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
D000596:Amino Acids; D002241:Carbohydrates; D002364:Caseins; D005732:Gangliosides; D006001:Glycoconjugates; D007797:Laminin; D037102:Lectins; D009844:Oligosaccharides; D010940:Plant Proteins; D012710:Serum Albumin, Bovine; D019158:N-Acetylneuraminic Acid