The contribution of tandem repeat number to the O-glycosylation of mucins

Howard Scott Silverman, Mark Sutton-Smith, Kimberly McDermott, Paul Heal, Shih Hsing Leir, Howard R. Morris, Michael A. Hollingsworth, Anne Dell, Ann Harris

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

The serine- and threonine-rich tandem repeat (TR) units that make up the characteristic feature of mucin glycoproteins are often polymorphic with substantial genetic variation in TR number. The precise effect of TR number on O-glycosylation is not fully understood, although the TR number of several mucins may be associated with apparent susceptibility to certain human diseases. To evaluate the contribution of TR number to O-glycosylation, we generated a series of chimeric mucins carrying increasing numbers of TR units from the MUC5B mucin in the context of an epitope-tagged MUC1 mucin backbone. These mucins were expressed in Caco2 colon carcinoma cell clones and purified by immunoprecipitation. O-Glycosylation was investigated by western blotting with antibodies to known carbohydrate structures and by fast atom bombardment-mass spectrometry. Additional carbohydrate epitopes; were detected with antibodies on chimeric mucins with a higher TR number in comparison to those with fewer TRs. Using mass spectrometry, higher-molecular-weight glycans were detected more frequently on the mucins with extended TRs compared to those with fewer TRs. However no novel carbohydrate structures were seen, suggesting that TR number does not affect the specificity of O-glycosylation.

Original languageEnglish (US)
Pages (from-to)265-277
Number of pages13
JournalGlycobiology
Volume13
Issue number4
DOIs
StatePublished - Apr 1 2003
Externally publishedYes

Fingerprint

Glycosylation
Tandem Repeat Sequences
Mucins
Carbohydrates
Mass spectrometry
Epitopes
Clone cells
Fast Atom Bombardment Mass Spectrometry
Antibodies
Threonine
Immunoprecipitation
Serine
Polysaccharides
Mass Spectrometry
Glycoproteins
Colon
Clone Cells
Molecular Weight
Western Blotting
Molecular weight

Keywords

  • MUC5B
  • Mucin tandem repeat
  • O-glycosylation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Scott Silverman, H., Sutton-Smith, M., McDermott, K., Heal, P., Leir, S. H., Morris, H. R., ... Harris, A. (2003). The contribution of tandem repeat number to the O-glycosylation of mucins. Glycobiology, 13(4), 265-277. https://doi.org/10.1093/glycob/cwg028

The contribution of tandem repeat number to the O-glycosylation of mucins. / Scott Silverman, Howard; Sutton-Smith, Mark; McDermott, Kimberly; Heal, Paul; Leir, Shih Hsing; Morris, Howard R.; Hollingsworth, Michael A.; Dell, Anne; Harris, Ann.

In: Glycobiology, Vol. 13, No. 4, 01.04.2003, p. 265-277.

Research output: Contribution to journalArticle

Scott Silverman, H, Sutton-Smith, M, McDermott, K, Heal, P, Leir, SH, Morris, HR, Hollingsworth, MA, Dell, A & Harris, A 2003, 'The contribution of tandem repeat number to the O-glycosylation of mucins', Glycobiology, vol. 13, no. 4, pp. 265-277. https://doi.org/10.1093/glycob/cwg028
Scott Silverman H, Sutton-Smith M, McDermott K, Heal P, Leir SH, Morris HR et al. The contribution of tandem repeat number to the O-glycosylation of mucins. Glycobiology. 2003 Apr 1;13(4):265-277. https://doi.org/10.1093/glycob/cwg028
Scott Silverman, Howard ; Sutton-Smith, Mark ; McDermott, Kimberly ; Heal, Paul ; Leir, Shih Hsing ; Morris, Howard R. ; Hollingsworth, Michael A. ; Dell, Anne ; Harris, Ann. / The contribution of tandem repeat number to the O-glycosylation of mucins. In: Glycobiology. 2003 ; Vol. 13, No. 4. pp. 265-277.
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