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Plakophilin 1 but not plakophilin 3 regulates desmoglein clustering

  • Michael Fuchs
  • Marco Foresti
  • Mariya Y. Radeva
  • Daniela Kugelmann
  • Rene Keil
  • Mechthild Hatzfeld
  • Volker Spindler
  • Jens WaschkeEmail author
  • Franziska VielmuthEmail author
Original Article
  • 63 Downloads

Abstract

Plakophilins (Pkp) are desmosomal plaque proteins crucial for desmosomal adhesion and participate in the regulation of desmosomal turnover and signaling. However, direct evidence that Pkps regulate clustering and molecular binding properties of desmosomal cadherins is missing. Here, keratinocytes lacking either Pkp1 or 3 in comparison to wild type (wt) keratinocytes were characterized with regard to their desmoglein (Dsg) 1- and 3-binding properties and their capability to induce Dsg3 clustering. As revealed by atomic force microscopy (AFM), both Pkp-deficient keratinocyte cell lines showed reduced membrane availability and binding frequency of Dsg1 and 3 at cell borders. Extracellular crosslinking and AFM cluster mapping demonstrated that Pkp1 but not Pkp3 is required for Dsg3 clustering. Accordingly, Dsg3 overexpression reconstituted cluster formation in Pkp3- but not Pkp1-deficient keratinocytes as shown by AFM and STED experiments. Taken together, these data demonstrate that both Pkp1 and 3 regulate Dsg membrane availability, whereas Pkp1 but not Pkp3 is required for Dsg3 clustering.

Keywords

Desmosome Cell adhesion Desmosomal clustering Atomic force microscopy STED 

Abbreviations

AFM

Atomic force microscopy

ARVC

Arrhythmogenic cardiomyopathy

DP

Desmoplakin

Dsc

Desmocollin

Dsg

Desmoglein

EC

Extracellular domains

FRAP

Fluorescence recovery after photobleaching

HaCaT

Cultured human keratinocytes

IF

Intermediate filament

MAPK

Mitogen-activated protein kinase

MKZ

Murine keratinocytes

PEG

Polyethylenglycol

Pg

Plakoglobin

Pkp

Plakophilins

PV

Pemphigus vulgaris

QI

Quantitative imaging

STED

Stimulated emission depletion

Sulfo-EGS

Ethylene glycolbis (sulfosuccinimidylsuccinate)

UFs

Unbinding forces

UP

Unbinding position

wt

Wild type

Notes

Acknowledgements

We thank Andrea Wehmeyer and Sabine Mühlsimer for excellent technical assistance and JPK instruments and Sunil Yeruva for constructive technical and scientific discussion. The project is funded by Else-Kröner-Fresenius-Stiftung 2016_AW157 to FV and JW and DFG SPP1782 to JW.

Author contributions

MFu, MF, MR, DK: methodology, data acquisition and analysis; FV and JW: funding acquisition, conceptualization and supervision; MFu, FV, JW: writing and editing, RK, MH, VS: methodology, review.

Compliance with ethical standards

Conflict of interest

The authors state that there was no conflict of interest.

Supplementary material

18_2019_3083_MOESM1_ESM.tif (24.9 mb)
Supplementary material 1 (TIFF 25518 kb)
18_2019_3083_MOESM2_ESM.tif (24.9 mb)
Supplementary material 2 (TIFF 25518 kb)
18_2019_3083_MOESM3_ESM.tif (24.9 mb)
Supplementary material 3 (TIFF 25518 kb)
18_2019_3083_MOESM4_ESM.docx (25 kb)
Supplementary material 4 (DOCX 24 kb)

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Michael Fuchs
    • 1
  • Marco Foresti
    • 1
  • Mariya Y. Radeva
    • 1
  • Daniela Kugelmann
    • 1
  • Rene Keil
    • 2
  • Mechthild Hatzfeld
    • 2
  • Volker Spindler
    • 3
  • Jens Waschke
    • 1
    Email author
  • Franziska Vielmuth
    • 1
    Email author
  1. 1.Faculty of Medicine, Institute of AnatomyLudwig-Maximilians-Universität MunichMunichGermany
  2. 2.Division of Pathobiochemistry, Institute of Molecular MedicineMartin-Luther-University Halle-WittenbergHalleGermany
  3. 3.Department of BiomedicineUniversity of BaselBaselSwitzerland

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