Purification and characterization of an N-acetyl-D-galactosamine-specific lectin from the edible mushroom Schizophyllum commune.
An N-acetyl-D-galactosamine (GalNAc)-specific lectin was purified from the edible mushroom, Schizophyllum commune, using affinity chromatography on a porcine stomach mucin (PSM)-Sepharose 4B column. Under reducing and non-reducing conditions, SDS-polyacrylamide gel electrophoresis gave a major band of 31.5 kDa. The Schizophyllum commune lectin (SCL) showed high affinity toward rat erythrocytes and the sugar inhibition assay exhibited its sugar specificity highly toward lactose and N-acetyl-D-galactosamine. It was stable at 55 degrees C for 30 min and at pH 3-10 for 18-h test. The lectin was shown to be a glycoprotein with cytotoxic activity against human epidermoid carcinoma cells. The N-terminus of SCL was blocked but amino acid sequences of internal tryptic peptides showed moderately sequence similarities with some other fungal and plant lectins. Crystals of SCL were obtained by the sitting drop vapour-diffusion method using polyethylene glycol 8000 as the precipitant, and gave an X-ray diffraction pattern to approximately 3.8 angstroms resolution.
Chumkhunthod
Podjana
P
School of Microbiology, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand. podjana@gmail.com
Rodtong
Sureelak
S
Lambert
Stan J
SJ
Fordham-Skelton
Anthony P
AP
Rizkallah
Pierre J
PJ
Wilkinson
Mark C
MC
Reynolds
Colin D
CD
D000116:Acetylgalactosamine / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D003460:Crystallization; D018360:Crystallography, X-Ray; D015180:Electrophoresis, Gel, Two-Dimensional; D006385:Hemagglutination Inhibition Tests; D037102:Lectins / Q000302:isolation & purification* / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D011817:Rabbits; D051381:Rats; D012567:Schizophyllum / Q000737:chemistry*
D037102:Lectins; D000116:Acetylgalactosamine