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 inBiochimica et biophysica acta Vol. 1760; no. 3; pp. 326 - 332
Main Authors Chumkhunthod, Podjana, Rodtong, Sureelak, Lambert, Stan J., Fordham-Skelton, Anthony P., Rizkallah, Pierre J., Wilkinson, Mark C., Reynolds, Colin D.
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.03.2006
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ISSN0304-4165
0006-3002
1872-8006
DOI10.1016/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.
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
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  givenname: Sureelak
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  organization: School of Microbiology, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand
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  givenname: Anthony P.
  surname: Fordham-Skelton
  fullname: Fordham-Skelton, Anthony P.
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  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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  ident: 10.1016/j.bbagen.2006.01.015_bib43
  article-title: The carbohydrate-binding specificity of a highly toxic protein from Abrus pulchellus seeds
  publication-title: Mem. Inst. Oswaldo Cruz
  doi: 10.1590/S0074-02761999000200010
– year: 2001
  ident: 10.1016/j.bbagen.2006.01.015_bib8
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Snippet An 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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StartPage 326
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
URI https://dx.doi.org/10.1016/j.bbagen.2006.01.015
https://www.ncbi.nlm.nih.gov/pubmed/16507335
https://www.proquest.com/docview/67756868
Volume 1760
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