Purification and Characterization of a Human Pancreatic Adenocarcinoma Mucin

Khorrami, Ali M. and Choudhury, Anirban Roy and Andrianifahanana, Mahefatiana and Varshney, Grish C and Bhattacharyya, Sambhu N. and Hollingsworth, Michael A. and Kaufman, Bernard and Batra, S K (2002) Purification and Characterization of a Human Pancreatic Adenocarcinoma Mucin. Journal of Biochemistry, 131 (1). pp. 21-29.

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Abstract

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 andMUC4mRNA, and moderate levels ofMUC5ACandMUC5BmRNA. The amino acid composition of the purified mucin shows a high degree of similarity to the MUC4 core protein.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Oxford University Press.
Uncontrolled Keywords:	adenocarcinomaDU-PAN-2glycoproteinsMUC4mucinspancreatic neoplasms
Subjects:	Q Science > QD Chemistry
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	06 Feb 2012 18:17
Last Modified:	22 Jul 2015 10:30
URI:	http://crdd.osdd.net/open/id/eprint/898

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