The hinge region of the scaffolding protein of cell contacts, zonula occludens protein 1, regulates interacting with various signaling proteins

Manjot Singh Bal, Victor Castro, Jörg Piontek, Christine Rueckert, Juliane K. Walter, Aliaksei Shymanets, Barbara Kurig, Hannelore Haase, Bernd Nürnberg, Ingolf E. Blasig

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Zonula occludens protein 1 (ZO-1) is a ubiquitous scaffolding protein, but it is unknown why it functions in very different cellular contacts. We hypothesized that a specific segment, the unique hinge region, can be bound by very different regulatory proteins. Using surface plasmon resonance spectroscopy and binding assays to peptide libraries, we show, for the first time, that the hinge region directly interacts with disparate signal elements such as G-proteins alpha 12 and alpha i2, the regulator of G-protein signaling 5, multifunctional signaling protein ahnak1, and L-type Ca2+-channel beta-2-subunit. The novel binding proteins specifically bound to a coiled coil-helix predicted in the hinge region of ZO-1. The interactions were modulated by phosphorylation in the hinge helix. Activation of the G-proteins influenced their association to ZO-1. In colon cells, G alpha i2 and ZO-1 were associated, as shown by coimmunoprecipitation. After cotransfection in kidney cells, G alpha i2 barely colocalized with ZO-1; the colocalization coefficient was significantly increased when epinephrine activated G-protein signaling. In conclusion, proteins with different regulatory potential adhere to and influence cellular functions of ZO-proteins and the interactions can be modulated via its hinge region and/or the binding proteins.

Original languageEnglish (US)
Pages (from-to)934-945
Number of pages12
JournalJournal of Cellular Biochemistry
Volume113
Issue number3
DOIs
StatePublished - Mar 1 2012

Keywords

  • Cell-cell contacts
  • G-proteins
  • Peptide binding assays
  • Protein-protein interaction
  • Regulatory proteins
  • Surface plasmon resonance spectroscopy

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Bal, M. S., Castro, V., Piontek, J., Rueckert, C., Walter, J. K., Shymanets, A., Kurig, B., Haase, H., Nürnberg, B., & Blasig, I. E. (2012). The hinge region of the scaffolding protein of cell contacts, zonula occludens protein 1, regulates interacting with various signaling proteins. Journal of Cellular Biochemistry, 113(3), 934-945. https://doi.org/10.1002/jcb.23422