May 2007
Volume 48, Issue 13
ARVO Annual Meeting Abstract  |   May 2007
Endogenous OA1 Is Distributed to the Apical Surface in RPE
Author Affiliations & Notes
  • R. C. Teeple
    Ophthalmology, University of Arizona, Tucson, Arizona
  • V. M. Lopez
    Ophthalmology, University of Arizona, Tucson, Arizona
  • B. S. McKay
    Ophthalmology, University of Arizona, Tucson, Arizona
  • Footnotes
    Commercial Relationships R.C. Teeple, None; V.M. Lopez, None; B.S. McKay, None.
  • Footnotes
    Support NIH EY014403 HIGHWIRE EXLINK_ID="48:5:2537:1" VALUE="EY014403" TYPEGUESS="GEN" /HIGHWIRE
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 2537. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      R. C. Teeple, V. M. Lopez, B. S. McKay; Endogenous OA1 Is Distributed to the Apical Surface in RPE. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2537.

      Download citation file:

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

  • Supplements

Purpose:: To determine the distribution of the endogenous OA1 protein in human RPE cells.

Methods:: Pairs of human donor eyes (N=8) were dissected to yield an eye cup with the apical surface of the RPE exposed. The cell surface was biotinylated using 1.0 mg/ml NHS-LC-Biotin in NaHCO3 buffered saline (pH 7.4) twice for 30min at 4oC. The reaction was terminated with 150mM glycine, 20mM Tris (pH 7.45). RPE were harvested in buffer containing 1% Triton X-100, 0.1% CHAPS, 1.0 mg/ml BSA, 200mM NaCl, 25mM NaHPO4 (pH 7.45). Biotinylated proteins were captured using streptavidin conjugated agarose, then bound and unbound proteins were subjected to western blot analysis. OA1 was identified and, as a control for cytoplasmic proteins, we probed with antibodies against actin. Similarly, selective cell surface biotinylation was performed using cells grown on filter supports to limit access to either the apical or basal compartment.

Results:: In each of the eight pairs of human eyes we detected OA1 on the surface of the RPE. We also detected a significant presence of OA1 in the unbound fraction, indicating a cytoplasmic distribution for the protein as well. Our control blots detected actin in only the unbound fraction, verifying that cytoplasmic proteins were not biotinylated. Further, OA1 was selectively detected on the apical surface of polarized RPE cells.

Conclusions:: Our data suggests that endogenous OA1 is both a cell surface and cytoplasmic protein. OA1 in RPE is distributed in a polarized manner to the apical surface.

Keywords: retinal pigment epithelium • receptors • protein purification and characterization 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.