Purification and Characterization of a Human Pancreatic Adenocarcinoma Mucin.
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概要
- 論文の詳細を見る
Pancreatic mucins consist of core proteins that are decorated with carbohydrate structures. Previous studies have identified at least two physically distinct populations of mucins produced by a pancreatic adenocarcinoma cell line (HPAF); one is the MUC1 core protein, which includes an oligosaccharide structure identified by a monoclonal antibody (MAb) recognizing the DU-PAN-2 epitope. In this study, we purified and characterized a second mucin fraction, which also shows reactivity with the DU-PAN-2 antibody, but which has an amino acid composition that is not consistent with the MUC1 core protein. This new mucin was purified by ammonium sulfate precipitation, molecular sieve chromatography, and density gradient centrifugation. It eluted in the void volume of a Sepharose 4B column together with an associated low molecular weight protein, which could be further resolved. The mucin is highly polyanionic due to numerous sulfated and sialylated saccharide chains. Carbohydrate analyses of the purified mucin showed the presence of galactose, glucosamine, galactosamine, and sialic acid, but no mannose, glucose, or uronic acid. The purified and deglycosylated mucin shows no reactivity with anti-MUC1 apomucin antibody, but reacts with antiserum against deglycosylated tracheal mucins and antiserum against the MUC4 tandem repeat peptide. Analysis of mucin expression in HPAF cells revealed high levels of MUC1 and MUC4 mRNA, and moderate levels of MUC5AC and MUC5B mRNA. The amino acid composition of the purified mucin shows a high degree of similarity to the MUC4 core protein.
- 社団法人 日本生化学会の論文
著者
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Hollingsworth Michael
Eppley Institute For Research In Cancer And Allied Diseases University Of Nebraska Medical Center
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Batra Surinder
Department Of Biochemistry And Molecular Biology University Of Nebraska Medical Center
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Kaufman Bernard
Department Of Biochemistry Duke University Medical Center
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Bhattacharyya Sambhu
Department Of Clinical Investigation William Beaumont Army Medical Center
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Khorrami Ali
S. U. N. Y. Upstate Medical University
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Varshney Grish
Department Of Biochemistry And Molecular Biology University Of Nebraska Medical Center
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Andrianifahanana Mahefatiana
Department Of Biochemistry And Molecular Biology University Of Nebraska Medical Center
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Choudhury Amit
Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center
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Choudhury Amit
Department of Biochemistry and Molecular Biology
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