Evidence for ezrin-radixin-moesin-binding phosphoprotein 50 (EBP50) self-association through PDZ-PDZ interactions

L. Fouassier, C. C. Yun, J. G. Fitz, R. B. Doctor

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

Ezrin-radixin-moesin (ERM)-binding phosphoprotein 50 (EBP50) is a aversatile membrane-cytoskeleton linking protein that binds to the COOH-tail of specific integral membrane proteins through its two PDZ domains. These EBP50 binding interactions have been implicated in sequestering interactive sets of proteins into common microdomains, regulating the activity of interacting proteins, and modulating membrane protein trafficking. With only two PDZ domains, it is unclear how EBP50 forms multiprotein complexes. Other PDZ proteins increase their breadth and diversity of protein interactions through oligomerization. Hypothesizing that EBP50 self-associates to amplify its functional capacity, far-Western blotting of cholangiocyte epithelial cell proreins with EBP50 fusion protein revealed that EBP50 binds to a 50-kDa protein. Far-Western blotting of EBP50 isolated by two-dimensional gel electrophoresis or immunoprecipitation demonstrates that the 50-kDa binding partner is itself EBP50. Further, co-transfection/co-precipitation studies show the self-association can occur in an intracellular environment. In vitro analysis of the EBP50-EBP50 binding interaction indicates it is both saturable and of relatively high affinity. Analysis of truncated EBP50 proteins indicates EBP50 self-association is mediated through its PDZ domains. The ability to self-associate provides a mechanism for EBP50 to expand its capacity to form multiprotein complexes and regulate membrane transport events.

Original languageEnglish (US)
Pages (from-to)25039-25045
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number32
DOIs
StatePublished - Aug 11 2000

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Phosphoproteins
Association reactions
PDZ Domains
Proteins
Far-Western Blotting
Multiprotein Complexes
ezrin
moesin
radixin
Membrane Proteins
Membranes
Oligomerization
Electrophoresis, Gel, Two-Dimensional
Protein Transport
Coprecipitation
Electrophoresis
Cytoskeleton
Immunoprecipitation
Transfection
Fusion reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Evidence for ezrin-radixin-moesin-binding phosphoprotein 50 (EBP50) self-association through PDZ-PDZ interactions. / Fouassier, L.; Yun, C. C.; Fitz, J. G.; Doctor, R. B.

In: Journal of Biological Chemistry, Vol. 275, No. 32, 11.08.2000, p. 25039-25045.

Research output: Contribution to journalArticle

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