Dimerization of the scaffolding protein ZO-1 through the second PDZ domain

Darkhan I. Utepbergenov, Alan S. Fanning, James M. Anderson

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

The tight junction protein ZO-1 is known to link the transmembrane proteins occludin, claudins, and JAMs to many cytoplasmic proteins and the actin cytoskeleton. Although specific roles for ZO-1 at the tight junction are unknown, it is widely assumed that ZO-1, together with its homologs ZO-2 and ZO-3, serves as a platform to scaffold various transmembrane and cytoplasmic tight junction proteins. Thus the manner in which the zonula occludens (ZO) proteins multimerize has implications for the protein networks they can coordinate. The purpose of our study was to determine whether ZO-1 forms homodimers and to determine the protein interaction region. Using laser light scattering and analytical centrifugation, we show that protein sequences corresponding to the NH2-terminal half of ZO-1 form stable homodimers with a submicromolar equilibrium dissociation constant. Analysis of the molecular weight of different truncated forms of ZO-1 revealed that the second PDZ domain is both necessary and sufficient for dimerization. This interaction does not use the β-finger motif described for other PDZ dimers. Furthermore, ZO-1 does not dimerize via an Src homology 3 to Guk domain interaction as was demonstrated previously for MAGUKs, like PSD-95. Results from immunoprecipitation experiments with polarized Madin-Darby canine kidney epithelial cells stably transfected with full-length GFP-ZO-1 indicate that a substantial portion of ZO-1 forms homodimers in vivo. As described previously, ZO-1 also forms heterodimers with ZO-2 and ZO-3. We conclude that the dimerization of ZO proteins is unlike that of other MAGUKs and that the previously unrecognized ZO-1 homodimers may allow formation of protein networks distinct from those of heterodimers with ZO-2 and ZO-3.

Original languageEnglish
Pages (from-to)24671-24677
Number of pages7
JournalJournal of Biological Chemistry
Volume281
Issue number34
DOIs
Publication statusPublished - Aug 25 2006
Externally publishedYes

Fingerprint

Zonula Occludens-1 Protein
PDZ Domains
Dimerization
Tight Junctions
Zonula Occludens Proteins
Tight Junction Proteins
Proteins
Claudins
Occludin
Centrifugation
Scaffolds
Dimers
Light scattering
Actins
Molecular weight
Lasers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Dimerization of the scaffolding protein ZO-1 through the second PDZ domain. / Utepbergenov, Darkhan I.; Fanning, Alan S.; Anderson, James M.

In: Journal of Biological Chemistry, Vol. 281, No. 34, 25.08.2006, p. 24671-24677.

Research output: Contribution to journalArticle

Utepbergenov, Darkhan I. ; Fanning, Alan S. ; Anderson, James M. / Dimerization of the scaffolding protein ZO-1 through the second PDZ domain. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 34. pp. 24671-24677.
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