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Purification and Characterization of a Human Pancreatic Adenocarcinoma Mucin

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Purification and Characterization of a Human Pancreatic Adenocarcinoma Mucin
 
Creator Khorrami, Ali M.
Choudhury, Anirban Roy
Andrianifahanana, Mahefatiana
Varshney, Grish C
Bhattacharyya, Sambhu N.
Hollingsworth, Michael A.
Kaufman, Bernard
Batra, S K
 
Subject QD Chemistry
 
Description 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.
 
Publisher Oxford University Press
 
Date 2002
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://crdd.osdd.net/open/898/1/varshney2002.1.pdf
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.
 
Relation http://jb.oxfordjournals.org/content/131/1/21.abstract
http://crdd.osdd.net/open/898/