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¹ Molecular Cell Biology Unit, Department for Molecular Biomedical Research, Flanders Interuniversity Institute for Biotechnology (VIB)-Ghent University, B-9000 Ghent, Belgium
² Molecular Biology Group of the Medical Faculty, Institute of Physiological Chemistry, University of Halle, 06097 Halle/Saale, Germany
³ Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611
⁴ Department of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611
⁵ The Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611

Plakophilin 3 (PKP3) is a recently described armadillo protein of the desmosomal plaque, which is synthesized in simple and stratified epithelia. We investigated the localization pattern of endogenous and exogenous PKP3 and fragments thereof. The desmosomal binding properties of PKP3 were determined using yeast two-hybrid, coimmunoprecipitation and colocalization experiments. To this end, novel mouse anti-PKP3 mAbs were generated. We found that PKP3 binds all three desmogleins, desmocollin (Dsc) 3a and -3b, and possibly also Dsc1a and -2a. As such, this is the first protein interaction ever observed with a Dsc-b isoform. Moreover, we determined that PKP3 interacts with plakoglobin, desmoplakin (DP) and the epithelial keratin 18. Evidence was found for the presence of at least two DP–PKP3 interaction sites. This finding might explain how lateral DP–PKP interactions are established in the upper layers of stratified epithelia, increasing the size of the desmosome and the number of anchoring points available for keratins. Together, these results show that PKP3, whose epithelial and epidermal desmosomal expression pattern and protein interaction repertoire are broader than those of PKP1 and -2, is a unique multiprotein binding element in the basic architecture of a vast majority of epithelial desmosomes.

Key Words: armadillo; cell adhesion; desmosomes; protein interaction; two-hybrid system

^* Abbreviations used in this paper: CBS, catenin-binding segment; CK18, cytokeratin 18; CoIP, coimmunoprecipitation; DP, desmoplakin; DPNTP, desmoplakin amino-terminal fragment; DPNTPmut, mutated desmoplakin amino-terminal fragment; Dsc, desmocollin; Dsg, desmoglein; IA, intracellular anchor domain; Pg, plakoglobin; PKP, plakophilin.

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