May 2004
Volume 45, Issue 13
ARVO Annual Meeting Abstract  |   May 2004
Predicted and measured disorder in the C–terminal domain of peripherin/rds.
Author Affiliations & Notes
  • A.F. X. Goldberg
    Eye Research Institute, Oakland University, Rochester, MI
  • L.M. Ritter
    Eye Research Institute, Oakland University, Rochester, MI
  • T. Arakawa
    Alliance Protein Laboratories, Thousand Oaks, CA
  • Footnotes
    Commercial Relationships  A.F.X. Goldberg, None; L.M. Ritter, None; T. Arakawa, None.
  • Footnotes
    Support  EY013246
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 2435. doi:
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    • Get Citation

      A.F. X. Goldberg, L.M. Ritter, T. Arakawa; Predicted and measured disorder in the C–terminal domain of peripherin/rds. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2435.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract: : Purpose: Elaboration of vertebrate photoreceptor outer segments (OSs) requires peripherin/rds, a tetrameric integral membrane protein that results in a surprising variety of progressive retinal degenerations when defective. This protein acts as an essential structural element for OS architecture in a manner that is not currently understood. In contrast to findings for the structurally important EC2 extracellular domain, a high degree of intrinsic disorder is predicted for the peripherin/rds C–terminus, both by secondary structure and PONDR VL–XT analyses; this observation suggests that this region may possess an underlying conformational flexibility. Methods: We have characterized the protein’s hydrophilic C–terminal domain by expressing and purifying a recombinant fusion protein from E. coli. Milligram quantities of the 63 C–terminal amino acids of bovine peripherin/rds could be liberated and affinity–purified from a GST header by proteolysis with a site–specific protease. Results: At all protein concentrations examined, this domain (CTER) behaved as a monomer in aqueous solution, as determined by analytical ultracentrifugation AUC. Despite its high expression levels and solubility, a variety of approaches, including: intrinsic tryptophan fluorescence, size–exclusion chromatography, and far ultraviolet circular dichroism, indicate that CTER lacks significant tertiary structure and possesses limited secondary structure. Conclusions: Conformational flexibility predicted for the peripherin/rds C–terminus has been documented experimentally for a recombinant protein produced in E. coli. Current studies are directed at determining whether intrinsic disorder plays a role in situ for peripherin/rds structure and function.

Keywords: photoreceptors • protein structure/function • proteins encoded by disease genes 

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