Purification and characterization of a human pancreatic adenocarcinoma mucin

Ali M. Khorrami, Amit Choudhury, Mahefatiana Andrianifahanana, Grish C. Varshney, Sambhu N. Bhattacharyya, Michael A. Hollingsworth, Bernard Kaufman, Surinder K. Batra

Research output: Contribution to journalArticle

15 Scopus citations

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 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.

Original languageEnglish (US)
Pages (from-to)21-29
Number of pages9
JournalJournal of Biochemistry
Volume131
Issue number1
DOIs
StatePublished - 2002

Keywords

  • Adenocarcinoma
  • DU-PAN-2
  • Glycoproteins
  • MUC4
  • Mucins
  • Pancreatic neoplasms

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Khorrami, A. M., Choudhury, A., Andrianifahanana, M., Varshney, G. C., Bhattacharyya, S. N., Hollingsworth, M. A., Kaufman, B., & Batra, S. K. (2002). Purification and characterization of a human pancreatic adenocarcinoma mucin. Journal of Biochemistry, 131(1), 21-29. https://doi.org/10.1093/oxfordjournals.jbchem.a003073