SUMOylation of claudin‐2

• DOI: 10.1111/j.1749-6632.2012.06541.x
• Publication Date: 2012-06-26T00:47:08Z
• Authors: Christina M. Van ...; Laura L. Mitic; James M. Anderson
• Keywords: Claudin; PDZ domain; Immunoprecipitation; RING finger domain

The C ‐terminal cytoplasmic tails of claudins are likely sites for interaction with proteins that regulate their function. We performed a yeast two‐hybrid screen the tail human claudin‐2 against kidney cDNA library and identified interactions PDZ3 domain ZO‐2 as well ubiquitin‐conjugating enzyme E2I (SUMO ligase‐1) E3 SUMO‐protein ligase PIAS; first is predicted interaction, while latter two novel suggest substrate SUMOylation. Using an in vitro SUMOylation assay, we K218 conjugation site on claudin‐2; mutation lysine to arginine blocked Stable expression inducible GFP‐SUMO‐1 MDCK cells resulted decreased levels protein by immunoblot membrane immunofluorescence microscopy. conclude cellular may be modulated SUMOylation, warranting further investigation pathways this modification vivo .