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In vitro evidence that peptides derived from the candidate tumor suppressor gene Esophageal Cancer‐Related Gene 4 (Ecrg4) internalize into cells through the innate immunity receptor complex
Author(s) -
Podvin Sonia,
Dang Xitong,
Kurabi Arwa,
Coimbra Raul,
Eliceiri Brian,
Baird Andrew
Publication year - 2012
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.26.1_supplement.998.2
Subject(s) - hek 293 cells , internalization , innate immune system , biology , receptor , insulin like growth factor 2 receptor , acquired immune system , microbiology and biotechnology , immune system , immunology , genetics , growth factor , insulin like growth factor 1 receptor
In light of data implicating the secreted precursor protein and candidate tumor suppressor Ecrg4 in the inflammatory response, we explored the possibility that predicted ligand domains in its primary sequence can target the innate immunity receptor complex. Predicted fragments of Ecrg4 were displayed on M13 phage and tested for internalization into cultured human embryonic kidney (HEK) cells that normally express low levels of the innate immunity receptor complex and HEK cells that stably overexpress. The receptor‐mediated internalized phage was recovered and phage DNA quantified by PCR. Internalized Ecrg4 targeted phage was compared to untargeted phage or epidermal growth factor (EGF) targeted phage that internalizes through the EGF receptor. Three peptide domains that derive from the Ecrg4 precursor internalized into HEK cells. Two of these, one at the amino terminus and one at the carboxyl terminus internalized 13 to 16 fold higher in HEK cells that overexpress TLR4/MD2/CD14 compared to wild type HEK cells. Untargeted phage internalization was low and EGF‐targeted phage internalization was high, but neither was different between the two cell lines. These data suggest that predicted Ecrg4 peptides derived from the secreted precursor protein can internalize into target cells through the innate immunity receptor complex. Supported by a Mentored Young Investigator award from the Hydrocephalus Association