Chemical and physicochemical characterization of the sialic acid-specific lectin from Cepaea hortensis.
A sialic acid-specific lectin was isolated from the albumin glands of the garden snail Cepaea hortensis by affinity chromatography on fetuin-Sepharose following gel filtration on Superdex 200. The purified native lectin showed a molecular mass of about 95 kDa by gel filtration and 100 kDa by SDS electrophoresis. It was cleaved by boiling in buffer containing SDS in three serological identical bands corresponding to molecular masses of about 24, 20 and 16 kDa, respectively. From these three fragments, only the 24- and the 20-kDa bands were found to be glycosylated. Only the three sugars mannose, galactose and N-acetylglucosamine could be detected in a molar ratio of 3:8.6:2. The oligosaccharide moieties seem to be N- and partially O-glycosidic bound. Isoelectric focusing (IEF) of the purified lectin revealed a heterogeneous pattern with bands in the pH range of 4.3-5.0. Isolated bands of different isoelectric points showed in SDS electrophoresis the same three fragments with molecular masses of 24, 20 or 16 kDa. The heterogeneity of the lectin was revealed either by IEF or amino acid sequencing of internal tryptic peptides.
Gerlach
Dieter
D
Friedrich-Schiller-University of Jena, Institute of Medical Microbiology, Semmelweisstr. 4, Germany. dieter.gerlach@med.uni-jena.de
Wagner
Manfred
M
Schlott
Bernhard
B
Zähringer
Ulrich
U
Schmidt
Karl-Herman
KH
D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006384:Hemagglutination; D007525:Isoelectric Focusing; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification; D019158:N-Acetylneuraminic Acid; D012685:Sepharose; D012908:Snails / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D037102:Lectins; C045831:agglutinin-I, cepaea hortensis; D012685:Sepharose; D019158:N-Acetylneuraminic Acid