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

被引:19
作者
Battacharya, G [1 ]
Cosgrove, D [1 ]
机构
[1] Boys Town Natl Res Hosp, Usher Syndrome Ctr, Omaha, NE 68131 USA
关键词
D O I
10.1021/bi050245u
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:11518 / 11524
页数:7
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