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...
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Published in | Biochimica et biophysica acta Vol. 1760; no. 3; pp. 326 - 332 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Netherlands
Elsevier B.V
01.03.2006
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Subjects | |
Online Access | Get full text |
ISSN | 0304-4165 0006-3002 1872-8006 |
DOI | 10.1016/j.bbagen.2006.01.015 |
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Abstract | 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 °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 Å resolution. |
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AbstractList | 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.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. 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 °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 Å resolution. 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. |
Author | Rodtong, Sureelak Wilkinson, Mark C. Lambert, Stan J. Reynolds, Colin D. Fordham-Skelton, Anthony P. Rizkallah, Pierre J. Chumkhunthod, Podjana |
Author_xml | – sequence: 1 givenname: Podjana surname: Chumkhunthod fullname: Chumkhunthod, Podjana email: podjana@gmail.com organization: School of Microbiology, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand – sequence: 2 givenname: Sureelak surname: Rodtong fullname: Rodtong, Sureelak organization: School of Microbiology, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand – sequence: 3 givenname: Stan J. surname: Lambert fullname: Lambert, Stan J. organization: School of Biomolecular Sciences, Max Perutz Building, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, England – sequence: 4 givenname: Anthony P. surname: Fordham-Skelton fullname: Fordham-Skelton, Anthony P. organization: CCLRC Daresbury Laboratory, Daresbury, Warrington, Cheshire WA4 4AD, England – sequence: 5 givenname: Pierre J. surname: Rizkallah fullname: Rizkallah, Pierre J. organization: CCLRC Daresbury Laboratory, Daresbury, Warrington, Cheshire WA4 4AD, England – sequence: 6 givenname: Mark C. surname: Wilkinson fullname: Wilkinson, Mark C. organization: School of Biological Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, England – sequence: 7 givenname: Colin D. surname: Reynolds fullname: Reynolds, Colin D. organization: School of Biomolecular Sciences, Max Perutz Building, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, England |
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Keywords | N-acetyl- d-galactosamine-specific lectin X-ray diffraction Lectin Schizophyllum commune Edible mushroom Lectin crystal |
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N-acetyl-
d-galactosamine (GalNAc)-specific lectin was purified from the edible mushroom,
Schizophyllum commune, using affinity chromatography on a porcine... An N-acetyl-D-galactosamine (GalNAc)-specific lectin was purified from the edible mushroom, Schizophyllum commune, using affinity chromatography on a porcine... |
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SubjectTerms | Acetylgalactosamine - chemistry Amino Acid Sequence Animals Chromatography, Affinity Crystallization Crystallography, X-Ray Edible mushroom Electrophoresis, Gel, Two-Dimensional Hemagglutination Inhibition Tests Lectin Lectin crystal Lectins - isolation & purification Lectins - pharmacology Microbial Sensitivity Tests N-acetyl- d-galactosamine-specific lectin Rabbits Rats Schizophyllum - chemistry Schizophyllum commune X-ray diffraction |
Title | Purification and characterization of an N-acetyl- d-galactosamine-specific lectin from the edible mushroom Schizophyllum commune |
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