Characterization of recombinant β subunit of human MUC4 mucin (rMUC4β)

Prakash G. Kshirsagar, Mansi Gulati, Wade M. Junker, Abhijit Aithal, Gaelle Spagnol, Srustidhar Das, Kavita Mallya, Shailendra K. Gautam, Sushil Kumar, Paul Sorgen, Krishan K. Pandey, Surinder K. Batra, Maneesh Jain

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


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.

Original languageEnglish (US)
Article number23730
JournalScientific reports
Issue number1
StatePublished - Dec 2021

ASJC Scopus subject areas

  • General


Dive into the research topics of 'Characterization of recombinant β subunit of human MUC4 mucin (rMUC4β)'. Together they form a unique fingerprint.

Cite this