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Glycosylation of Human MUC1 Transcript Variant 2 Expressed in HeLa Cells
Author(s) -
Warren Janine,
Piefer Andrew
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.571.29
Subject(s) - glycosylation , muc1 , immunoprecipitation , gene isoform , tandem repeat , epitope , recombinant dna , glycan , blot , microbiology and biotechnology , n linked glycosylation , biology , hela , biochemistry , chemistry , antigen , gene , glycoprotein , in vitro , mucin , genetics , genome
The purpose of this project is to study the localization and glycosylation of MUC1 a potentially important antigen found on tumor cells. Naïve and carcinogenic cells produce this protein, but there are very distinct differences between MUC1 isoforms found on these cells. MUC1 has a 20 amino acid tandem repeat where the bulk of O‐linked glycosylation occurs. There are five spots in each tandem repeat (TR) for glycosylation, but the typical MUC1 isoform only fills about 2.5 glycosylations per TR. Recognition of an altered cancer specific MUC1 glycosylation may lead to an effective target for anti‐cancer therapies. A recombinant, epitope‐tagged Human MUC1 transcript variant 2 cDNA will be transiently expressed in HeLa cells to study protein glycosylation. MUC1 will be isolated via immunoprecipitation and subjected to glycosidase treatment to determine presence of sugars. SDS‐PAGE and western blotting will be used to detect MUC1. We hypothesize that the glycosylation of recombinant MUC1 produced in HeLa cells will be similar to the glycosylation patterns found on MUC1 present in epithelial tumors. This project was funded by the Hartwick College Department of Chemistry.