May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
NO-Induced Lipofuscin Acumulation Was Caused by Inhibition of Cathepsin S
Author Affiliations & Notes
  • E. Sugano
    Ophthalmology, Tohoku Univ School of Medicine, Sendai, Japan
  • H. Tomita
    Ophthalmology, Tohoku Univ School of Medicine, Sendai, Japan
  • T. Abe
    Ophthalmology, Tohoku Univ School of Medicine, Sendai, Japan
  • H. Taira
    Ophthalmology, Tohoku Univ School of Medicine, Sendai, Japan
  • M. Tamai
    Ophthalmology, Tohoku Univ School of Medicine, Sendai, Japan
  • Footnotes
    Commercial Relationships  E. Sugano, None; H. Tomita, None; T. Abe, None; H. Taira, None; M. Tamai, None.
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 1624. doi:
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    • Get Citation

      E. Sugano, H. Tomita, T. Abe, H. Taira, M. Tamai; NO-Induced Lipofuscin Acumulation Was Caused by Inhibition of Cathepsin S . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1624.

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

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Abstract

Abstract: : Purpose:To investigate effects of Nitric oxide (NO) on the phagocytosis in RPE cells with the correlation of cathepsin S (Cat S) activity.Methods:RPE cells were isolated from 10-week-old Long evans rats. The RPE cells were cultured in the medium containing NO donor (S-nitroso-N-acetylpenicillamine,SNAP) with or without NO scavenger (Hydroxocobalamin) from 72hrs before the measurement of the phagocytic activities. Phagocytic activities of ROS were evaluated by a flow cytometric analysis and the activity of Cat S was measured by using a fluorogenic substrate. We also measured the activity of purified bovine spleen Cat S under the presence of SNAP. The Labeled or unlabeled ROS was used for monitoring the ingestion or digestion of ROS in RPE cells, respectively. Results:SNAP inhibited the phagocytic and Cat S activities of the RPE cells in a dose-dependent manner. Purified bovine spleen catS activity was also suppressed by co-incubation of SNAP in a dose dependent manner. These activities were partially recovered by the addition of NO scavenger in both RPE cells and purified Cat S. Conclusions: We demonstrated that NO donor inhibited the activity of catS and the phagosytosis of ROS. There were some reports about the direct S- nitrosylation of caspase catalytic cysteine residue by NO and Cat S is a cystein protease. The inhibitory effects of NO on phagocytosis may be due to S-nitrosylation which involves transfer of the NO group to the sulfhydryl of a cysteine residue.

Keywords: phagocytosis and killing • nitric oxide • enzymes/enzyme inhibitors 
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