May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Interaction of Secreted Myocilin and the Cell Surface Protein on NIH3T3 Cells
Author Affiliations & Notes
  • M. Minami
    National Institute of Sensory Organs, National Hospital Organization Tokyo Medical Center, Meguro-ku, Japan
  • T. Iwata
    National Institute of Sensory Organs, National Hospital Organization Tokyo Medical Center, Meguro-ku, Japan
  • Footnotes
    Commercial Relationships M. Minami, None; T. Iwata, None.
  • Footnotes
    Support Ministry of Health, Labour and Welfare of Japan
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 5921. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      M. Minami, T. Iwata; Interaction of Secreted Myocilin and the Cell Surface Protein on NIH3T3 Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):5921.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose:: Myocilin is known to be secreted into an aqueous humor. In this study, secreted myocilin was applied to various cultured cells to investigate whether specific interaction with cell surface protein may occur.

Methods:: Histidine-tagged recombinant myocilin was expressed in COS-1 cells and the culture medium containing the secreted myocilin was collected. This medium was added RGC-5, A549, and NIH3T3 cells and these cells were cultured for 72-96 hours. Cell surface distribution of myocilin was determined by immunostaining.

Results:: Immunofluorescence microscopic analysis revealed that secreted myocilin was bound specifically to cell surface of NIT3T3 cells. Localization of myocilin was dot-like pattern, and this interaction was dependent on cell density. Binding of myocilin to cell surface was not observed at the low density of 7.5 x 102/cm2, whereas myocilin could associate with NIH3T3 cells at the high density of 1.5 x 103/cm2. Recombinant myocilin and unknown cell surface protein(s) were cross-linked by membrane impermeable cross-linker (sulfosuccinimidylpropionate) followed by cell lysis and immunoprecipitation.

Conclusions:: Myocilin was shown to interact with cell surface in a specific manner. These results indicate that myocilin is capable of cell surface interaction. Currently, precipitated proteins are under investigation.

Keywords: cell-cell communication • trabecular meshwork • signal transduction 
×
×

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.

×