RGD Reference Report - The rat IgGFcgammaBP and Muc2 C-terminal domains and TFF3 in two intestinal mucus layers bind together by covalent interaction. - Rat Genome Database

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The rat IgGFcgammaBP and Muc2 C-terminal domains and TFF3 in two intestinal mucus layers bind together by covalent interaction.

Authors: Yu, H  He, Y  Zhang, X  Peng, Z  Yang, Y  Zhu, R  Bai, J  Tian, Y  Li, X  Chen, W  Fang, D  Wang, R 
Citation: Yu H, etal., PLoS One. 2011;6(5):e20334. doi: 10.1371/journal.pone.0020334. Epub 2011 May 23.
RGD ID: 7349366
Pubmed: PMID:21629776   (View Abstract at PubMed)
PMCID: PMC3100329   (View Article at PubMed Central)
DOI: DOI:10.1371/journal.pone.0020334   (Journal Full-text)

BACKGROUND: The secreted proteins from goblet cells compose the intestinal mucus. The aims of this study were to determine how they exist in two intestinal mucus layers. METHODOLOGY/PRINCIPAL FINDINGS: The intestinal mucosa was fixed with Carnoy solution and immunostained. Mucus from the loose layer, the firm layer was gently suctioned or scraped, respectively, lysed in SDS sample buffer with or without DTT, then subjected to the western blotting of rTFF3, rIgGFcgammaBP or rMuc2. The non-reduced or reduced soluble mucus samples in RIPA buffer were co-immunoprecipitated to investigate their possible interactions. Polyclonal antibodies for rTFF3, the rIgGFcgammaBP C-terminal domain and the rMuc2 C-terminal domain confirmed their localization in the mucus layer and in the mucus collected from the rat intestinal loose layer or firm layer in both western blot and immunoprecipitation experiments. A complex of rTFF3, which was approximately 250 kDa, and a monomer of 6 kDa were present in both layers of the intestinal mucus; rIgGFcgammaBP was present in the complex (250-280 kDa) under non-reducing conditions, but shifted to 164 kDa under reducing conditions in both of the layers. rMuc2 was found mainly in a complex of 214-270 kDa under non-reducing conditions, but it shifted to 140 kDa under reducing conditions. The co-immunoprecipitation experiments showed that binding occurs among rTFF3, rIgGFcgammaBP and rMuc2 in the RIPA buffer soluble intestinal mucus. Blocking the covalent interaction by 100 mM DTT in the RIPA buffer soluble intestinal mucus disassociated their binding. CONCLUSIONS/SIGNIFICANCE: Rat goblet cell-secreted TFF3, IgGFcgammaBP and Muc2, existing in the two intestinal mucus layers, are bound together by covalent interactions in the soluble fraction of intestinal mucus and form heteropolymers to be one of the biochemical mechanisms of composing the net-like structure of mucus.

Gene Ontology Annotations    Click to see Annotation Detail View

Cellular Component
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
mucus layer  IDA 7349366 RGD 

Molecular Function
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
protein binding  IPITff3 (Rattus norvegicus)7349366 RGD 
protein binding  IPIMuc2 (Rattus norvegicus)7349366 RGD 

Objects Annotated

Genes (Rattus norvegicus)
Muc2  (mucin 2, oligomeric mucus/gel-forming)
Tff3  (trefoil factor 3)

Additional Information