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Extraction and Verification of Mouse and Human Mucins from Tissue and Fecal Material

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Carbohydrate-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2657))

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Abstract

Gel-forming mucins are highly O-glycosylated polymeric glycoproteins which have critical roles in tissue protection from environmental insult. To understand their biochemical properties, these samples must be extracted and enriched from biological samples. Here we describe how to extract and semi-purify human and murine mucins from intestinal scrapings or fecal material. As mucins have high molecular weights, traditional gel electrophoresis methods are unable to effectively separate these glycoproteins for analysis. We describe the procedure for making composite sodium dodecyl sulfate urea agarose-polyacrylamide (SDS-UAgPAGE) gels, which allows for accurate verification and band separation of extracted mucins.

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References

  1. Michael AM, David JT, Jeffrey AW (2015) Chapter 14 – Mucins and mucus. In: Jiri M, Warren S, Michael WR, Brian LK, Hilde C, Bart NL (eds) Mucosal immunology, 4th edn. Academic, Boston, pp 231–250. https://doi.org/10.1016/B978-0-12-415847-4.00014-8

    Chapter  Google Scholar 

  2. Malin EVJ, Gunnar CH (2016) The mucins. In: Michael JHR (ed) Encyclopedia of immunobiology. Academic, Oxford, pp 381–388. https://doi.org/10.1016/B978-0-12-374279-7.02019-1

    Chapter  Google Scholar 

  3. Godl K, Johansson ME, Lidell ME, Morgelin M, Karlsson H, Olson FJ et al (2002) The N terminus of the MUC2 mucin forms trimers that are held together within a trypsin-resistant core fragment. J Biol Chem 277(49):47248–47256. https://doi.org/10.1074/jbc.M208483200

    Article  CAS  PubMed  Google Scholar 

  4. Carlstedt I, Herrmann A, Hovenberg H, Lindell G, Nordman H, Wickstrom C et al (1995) ‘Soluble’ and ‘insoluble’ mucins – identification of distinct populations. Biochem Soc Trans 23(4):845–851. https://doi.org/10.1042/bst0230845

    Article  CAS  PubMed  Google Scholar 

  5. Johansson ME, Hansson GC (2012) Analysis of assembly of secreted mucins. Methods Mol Biol 842:109–121. https://doi.org/10.1007/978-1-61779-513-8_6

    Article  CAS  PubMed  Google Scholar 

  6. Davies JR, Carlstedt I (2000) Isolation of large gel-forming mucins. Methods Mol Biol 125:3–13. https://doi.org/10.1385/1-59259-048-9:003

    Article  CAS  PubMed  Google Scholar 

  7. Herrmann A, Davies JR, Lindell G, Martensson S, Packer NH, Swallow DM et al (1999) Studies on the “insoluble” glycoprotein complex from human colon. Identification of reduction-insensitive MUC2 oligomers and C-terminal cleavage. J Biol Chem 274(22):15828–15836. https://doi.org/10.1074/jbc.274.22.15828

    Article  CAS  PubMed  Google Scholar 

  8. Tytgat KM, Swallow DM, Van Klinken BJ, Buller HA, Einerhand AW, Dekker J (1995) Unpredictable behaviour of mucins in SDS/polyacrylamide-gel electrophoresis. Biochem J 310(Pt 3):1053–1054. https://doi.org/10.1042/bj3101053

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Schulz BL, Packer NH, Karlsson NG (2002) Small-scale analysis of O-linked oligosaccharides from glycoproteins and mucins separated by gel electrophoresis. Anal Chem 74(23):6088–6097. https://doi.org/10.1021/ac025890a

    Article  CAS  PubMed  Google Scholar 

  10. Issa SM, Schulz BL, Packer NH, Karlsson NG (2011) Analysis of mucosal mucins separated by SDS-urea agarose polyacrylamide composite gel electrophoresis. Electrophoresis 32(24):3554–3563. https://doi.org/10.1002/elps.201100374

    Article  CAS  PubMed  Google Scholar 

  11. Thomsson KA, Hansson GC (2011) Identification and quantification of mucin expression. Methods Mol Biol 742:127–141. https://doi.org/10.1007/978-1-61779-120-8_8

    Article  CAS  PubMed  Google Scholar 

  12. Bergstrom K, Shan X, Casero D, Batushansky A, Lagishetty V, Jacobs JP et al (2020) Proximal colon-derived O-glycosylated mucus encapsulates and modulates the microbiota. Science 370(6515):467–472. https://doi.org/10.1126/science.aay7367

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Department of Biology, The University of British Columbia, Kelowna, BC, Canada

    Mackenzie Melvin, Noah Fancy, Darrek Kniffen & Kirk Bergstrom

Authors
  1. Mackenzie Melvin
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  2. Noah Fancy
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  3. Darrek Kniffen
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  4. Kirk Bergstrom
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Corresponding author

Correspondence to Kirk Bergstrom .

Editor information

Editors and Affiliations

  1. Agriculture and Agri-Food Canada, Lethbridge Research and Development Center, Lethbridge, AB, Canada

    D. Wade Abbott

  2. Department of Biochemistry, The University of British Columbia, Kelowna, BC, Canada

    Wesley F. Zandberg

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Melvin, M., Fancy, N., Kniffen, D., Bergstrom, K. (2023). Extraction and Verification of Mouse and Human Mucins from Tissue and Fecal Material. In: Abbott, D.W., Zandberg, W.F. (eds) Carbohydrate-Protein Interactions. Methods in Molecular Biology, vol 2657. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3151-5_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3151-5_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3150-8

  • Online ISBN: 978-1-0716-3151-5

  • eBook Packages: Springer Protocols

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