A domain-specific usherin/collagen IV interaction may be required for stable integration into the basement membrane superstructure

Gautam Bhattacharya, Raghu Kalluri, Dana J. Orten, William J. Kimberling, Dominic Cosgrove

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

Usherin is a basement membrane protein encoded by the gene associated with Usher syndrome type IIa, the most common deaf/blind disorder. This report demonstrates a specific interaction between type IV collagen and usherin in the basement membrane, with a 1:1 stoichiometry for binding. Genetic and biochemical approaches were used to explore the role of type IV collagen binding in usherin function. We demonstrate binding occurs between the LE domain of usherin and the 7S domain of type IV collagen. A purified fusion peptide comprising the first four LE modules was shown to compete with full-length recombinant usherin for type IV collagen binding. However, synonymous fusion peptides with single amino acid substitutions resulting from missense mutations that were known to cause Usher syndrome type IIa in humans, failed to compete. Only mutations in loop b of the LE domain abolished binding activity. Co-immunoprecipitation and western blot analysis of testicular basement membranes from the Alport mouse model show a 70% reduction in type IV collagen is associated with a similar reduction in usherin, suggesting the usherin/collagen (IV) interaction stabilizes usherin in the basement membrane. Thus, the domain-specific interaction between usherin and type IV collagen appears essential to usherin stability in vivo, and loss of this interaction may result in Usher pathology in humans.

Original languageEnglish (US)
Pages (from-to)233-242
Number of pages10
JournalJournal of cell science
Volume117
Issue number2
DOIs
StatePublished - Jan 15 2004

Keywords

  • Basement membrane
  • Collagen (IV)
  • Protein interaction
  • Usher syndrome
  • Usherin

ASJC Scopus subject areas

  • Cell Biology

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