Characterization of recombinant β subunit of human MUC4 mucin (rMUC4β).
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
09 12 2021
09 12 2021
Historique:
received:
13
08
2021
accepted:
03
11
2021
entrez:
10
12
2021
pubmed:
11
12
2021
medline:
1
2
2022
Statut:
epublish
Résumé
MUC4 is a transmembrane mucin expressed on various epithelial surfaces, including respiratory and gastrointestinal tracts, and helps in their lubrication and protection. MUC4 is also aberrantly overexpressed in various epithelial malignancies and functionally contributes to cancer development and progression. MUC4 is putatively cleaved at the GDPH site into a mucin-like α-subunit and a membrane-tethered growth factor-like β-subunit. Due to the presence of several functional domains, the characterization of MUC4β is critical for understanding MUC4 biology. We developed a method to produce and purify multi-milligram amounts of recombinant MUC4β (rMUC4β). Purified rMUC4β was characterized by Far-UV CD and I-TASSER-based protein structure prediction analyses, and its ability to interact with cellular proteins was determined by the affinity pull-down assay. Two of the three EGF-like domains exhibited typical β-fold, while the third EGF-like domain and vWD domain were predominantly random coils. We observed that rMUC4β physically interacts with Ezrin and EGFR family members. Overall, this study describes an efficient and simple strategy for the purification of biologically-active rMUC4β that can serve as a valuable reagent for a variety of biochemical and functional studies to elucidate MUC4 function and generating domain-specific antibodies and vaccines for cancer immunotherapy.
Identifiants
pubmed: 34887447
doi: 10.1038/s41598-021-02860-5
pii: 10.1038/s41598-021-02860-5
pmc: PMC8660890
doi:
Substances chimiques
MUC4 protein, human
0
Mucin-4
0
Protein Subunits
0
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
23730Subventions
Organisme : NCI NIH HHS
ID : R01 CA247471
Pays : United States
Organisme : NIDDK NIH HHS
ID : R44 DK117472
Pays : United States
Organisme : NCI NIH HHS
ID : P01 CA217798
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA195586
Pays : United States
Organisme : NCI NIH HHS
ID : U01 CA213862
Pays : United States
Informations de copyright
© 2021. The Author(s).
Références
AMB Express. 2018 Aug 22;8(1):135
pubmed: 30136189
FASEB J. 2008 Apr;22(4):966-81
pubmed: 18024835
Br J Cancer. 2007 Aug 6;97(3):345-57
pubmed: 17595659
Nat Methods. 2015 Jan;12(1):7-8
pubmed: 25549265
Eur J Biochem. 2002 Jan;269(1):2-12
pubmed: 11784292
Biochem J. 1999 Mar 1;338 ( Pt 2):325-33
pubmed: 10024507
Future Oncol. 2009 Dec;5(10):1631-40
pubmed: 20001800
Gene Ther. 2011 Feb;18(2):135-44
pubmed: 20668485
Clin Cancer Res. 2011 Jan 15;17(2):267-74
pubmed: 21059814
Protein Sci. 2003 Feb;12(2):384-8
pubmed: 12538903
Proteins. 2005 Jan 1;58(1):144-50
pubmed: 15476208
Nat Rev Gastroenterol Hepatol. 2013 Oct;10(10):607-20
pubmed: 23856888
Nat Rev Cancer. 2009 Dec;9(12):874-85
pubmed: 19935676
Invest Ophthalmol Vis Sci. 2007 Oct;48(10):4509-18
pubmed: 17898272
Semin Immunol. 2020 Feb;47:101391
pubmed: 31952903
Trends Biochem Sci. 2010 Apr;35(4):236-45
pubmed: 19913432
Cancer Metastasis Rev. 2019 Jun;38(1-2):223-236
pubmed: 30618016
J Transl Med. 2010 Jun 23;8:61
pubmed: 20569470
Expert Opin Med Diagn. 2008 Aug;2(8):891-910
pubmed: 23495864
Proc Natl Acad Sci U S A. 2015 Jun 16;112(24):E3095-103
pubmed: 26038575
Expert Opin Ther Targets. 2017 Jul;21(7):657-669
pubmed: 28460571
J Biosci Bioeng. 2005 Apr;99(4):303-10
pubmed: 16233795
Appl Environ Microbiol. 1999 Apr;65(4):1524-9
pubmed: 10103246
PLoS One. 2013;8(2):e54455
pubmed: 23408941
Biomolecules. 2014 Feb 20;4(1):235-51
pubmed: 24970214
PLoS One. 2018 Apr 30;13(4):e0193907
pubmed: 29708979
IUBMB Life. 2005 Jul;57(7):505-12
pubmed: 16081372
Nat Protoc. 2010 Apr;5(4):725-38
pubmed: 20360767
Cancer Res. 2007 Jan 15;67(2):433-6
pubmed: 17234748
Oncogene. 2010 May 20;29(20):2893-904
pubmed: 20348949
J Biomed Mater Res A. 2021 Jun;109(6):893-902
pubmed: 32776461
Br J Cancer. 2008 Aug 5;99(3):520-6
pubmed: 18665193
Sci Rep. 2019 Nov 13;9(1):16678
pubmed: 31723153
Curr Opin Biotechnol. 2002 Oct;13(5):418-23
pubmed: 12459331
Oncotarget. 2015 Aug 28;6(25):21085-99
pubmed: 26035354
Nat Rev Cancer. 2004 Jan;4(1):45-60
pubmed: 14681689
Mol Cancer Res. 2007 Apr;5(4):309-20
pubmed: 17406026
Bioinformatics. 2011 Jun 15;27(12):1630-6
pubmed: 21505036
Expert Opin Ther Targets. 2018 Aug;22(8):675-686
pubmed: 29999426
Genes Cancer. 2019 May;10(3-4):52-62
pubmed: 31258832
Cell Adh Migr. 2011 Mar-Apr;5(2):199-206
pubmed: 21343695
PLoS One. 2011;6(8):e23344
pubmed: 21886786
Nucleic Acids Res. 2015 Jul 1;43(W1):W174-81
pubmed: 25883148
Protein Expr Purif. 2010 Feb;69(2):159-67
pubmed: 19733669
Protein Expr Purif. 2006 Nov;50(1):49-57
pubmed: 16962338
Eur J Biochem. 2000 Jul;267(14):4536-44
pubmed: 10880978
Protein Expr Purif. 2000 Aug;19(3):419-24
pubmed: 10910733
Nucleic Acids Res. 2007;35(10):3375-82
pubmed: 17478507
Cancer Res. 2008 Nov 15;68(22):9231-8
pubmed: 19010895
Proteins. 2002 Nov 1;49(2):154-66
pubmed: 12210997
Cancer Biother Radiopharm. 2011 Jun;26(3):261-71
pubmed: 21728842
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W668-73
pubmed: 15215473
Anal Biochem. 1987 Nov 15;167(1):76-85
pubmed: 3434802
Anal Biochem. 2000 Dec 15;287(2):252-60
pubmed: 11112271
Bioresour Technol. 2018 Aug;261:176-181
pubmed: 29660658
J Cell Sci. 2014 Jan 15;127(Pt 2):267-75
pubmed: 24421310
PLoS One. 2012;7(2):e32232
pubmed: 22393391
Carcinogenesis. 2017 Jul 1;38(7):671-679
pubmed: 27838635
FASEB J. 2020 Feb;34(2):1939-1957
pubmed: 31908009
Cancer Res. 2008 Apr 1;68(7):2065-70
pubmed: 18381409
Nat Protoc. 2006;1(1):418-28
pubmed: 17406264