Skip to main content

Extraction of Mucins from the Mammalian Intestinal Tract

  • Protocol
  • First Online:
Mucins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2763))

  • 349 Accesses

Abstract

In the intestine, mucus covering the mucosa plays a critical role in maintaining gut homeostasis by protecting the mucosa from invasion by commensal bacteria. The gut mucus is composed primarily of MUC2 mucin secreted by goblet cells. MUC2 is highly O-glycosylated, and O-glycans are necessary for the function and polymer structure of MUC2. In addition, recent evidence revealed that several glycan modifications, such as sialylation and sulfation, confer resistance of mucins to proteolysis and affect the viscosity and lubricity of mucus. Therefore, characterizing glycan structures of mucins is required to understand their functions fully. In this chapter, we describe how to purify secreted mucins from the mammalian intestine for analysis of their glycan structures. This description includes the extraction of MUC2 mucin from the mucosal surface of the mouse colon and colon explants.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Okumura R, Takeda K (2018) Maintenance of intestinal homeostasis by mucosal barriers. Inflamm Regen 38:5

    Article  PubMed  PubMed Central  Google Scholar 

  2. McGuckin MA, Lindén SK, Sutton P et al (2011) Mucin dynamics and enteric pathogens. Nat Rev Microbiol 9:265–278

    Article  PubMed  Google Scholar 

  3. Johansson ME, Larsson JM, Hansson GC (2011) The two mucus layers of colon are organized by the MUC2 mucin, whereas the outer layer is a legislator of host-microbial interactions. Proc Natl Acad Sci U S A 108:4659–4665

    Article  PubMed  Google Scholar 

  4. Paone P, Cani PD (2020) Mucus barrier, mucins and gut microbiota: the expected slimy partners? Gut 69:2232–2243

    Article  PubMed  Google Scholar 

  5. Pickard JM, Chervonsky AV (2015) Intestinal fucose as a mediator of host-microbe symbiosis. J Immunol 194:5588–5593

    Article  PubMed  Google Scholar 

  6. Kudelka MR, Stowell SR, Cummings RD et al (2020) Intestinal epithelial glycosylation in homeostasis and gut microbiota interactions in IBD. Nat Rev Gastroenterol Hepatol 17:597–617

    Article  PubMed  PubMed Central  Google Scholar 

  7. Kufe DW (2009) Mucins in cancer: function, prognosis and therapy. Nat Rev Cancer 9:874–885

    Article  PubMed  PubMed Central  Google Scholar 

  8. Brockhausen I (2006) Mucin-type O-glycans in human colon and breast cancer: glycodynamics and functions. EMBO Rep 7:599–604

    Article  PubMed  PubMed Central  Google Scholar 

  9. Kesimer M, Sheehan JK (2012) Mass spectrometric analysis of mucin core proteins. Methods Mol Biol 842:67–79

    Article  PubMed  PubMed Central  Google Scholar 

  10. Davies JR, Wickström C, Thornton DJ (2012) Gel-forming and cell-associated mucins: preparation for structural and functional studies. Methods Mol Biol 842:27–47

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan: JP18K15187, Uehara Memorial Foundation and Takeda Science Foundation. We thank Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ryu Okumura .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Okamoto, S., Taniguchi, M., Okumura, R. (2024). Extraction of Mucins from the Mammalian Intestinal Tract. In: Kameyama, A. (eds) Mucins. Methods in Molecular Biology, vol 2763. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3670-1_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3670-1_6

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3669-5

  • Online ISBN: 978-1-0716-3670-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics