Evidence for functional importance of usherin/fibronectin interactions in retinal basement membranes

Gautam Bhattacharya, Dominic E Cosgrove

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Usher syndrome is a genetically heterogeneous disorder characterized by hearing loss with retinitis pigmentosa. Usher syndrome type IIa is the most common of the Usher syndromes, accounting for over half of all cases. The gene encodes a 180 kDa basement membrane glycoprotein called usherin. Here, we demonstrated a specific interaction between usherin and fibronectin in retinal basement membranes. This interaction was confirmed using biochemical, biophysical, and genetic approaches. Surface plasmon resonance assay confirmed that fibronectin binding to usherin is of high affinity and 1:1 stoichiometry. Using a fusion peptide-based co-immunoprecipitation approach, we show that binding to fibronectin occurs at the LE domain of usherin. Recombinant LE domain-specific peptides were engineered that contained single amino acid substitutions corresponding to missense mutations found in humans with Usher syndrome type IIa. Only mutations in loop d of the LE domain abolished the ability of the LE domain to co-immunoprecipitate fibronectin.

Original languageEnglish (US)
Pages (from-to)11518-11524
Number of pages7
JournalBiochemistry
Volume44
Issue number34
DOIs
StatePublished - Aug 30 2005

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Usher Syndromes
Fibronectins
Basement Membrane
Peptides
Retinitis Pigmentosa
Surface Plasmon Resonance
Membrane Glycoproteins
Audition
Surface plasmon resonance
Missense Mutation
Amino Acid Substitution
Hearing Loss
Immunoprecipitation
Stoichiometry
Molecular Biology
Assays
Substitution reactions
Fusion reactions
Genes
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Evidence for functional importance of usherin/fibronectin interactions in retinal basement membranes. / Bhattacharya, Gautam; Cosgrove, Dominic E.

In: Biochemistry, Vol. 44, No. 34, 30.08.2005, p. 11518-11524.

Research output: Contribution to journalArticle

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