IOVS
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

(Investigative Ophthalmology and Visual Science. 2003;44:3263-3271.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-1322
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yegorova, S.
Right arrow Articles by Lou, M. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yegorova, S.
Right arrow Articles by Lou, M. F.

Human Lens Thioredoxin: Molecular Cloning and Functional Characterization

Svitlana Yegorova,1 Aimin Liu,1 and Marjorie F. Lou1,2,3

1From the Departments of Veterinary and Biomedical Sciences and 2Biochemistry, University of Nebraska-Lincoln, Lincoln, Nebraska; and the 3Department of Ophthalmology, University of Nebraska Medical Center, Omaha, Nebraska.

PURPOSE. To molecularly clone the human lens thioredoxin (TXN) gene, characterize the recombinant protein (rTrx1) and study the regulation expression of thioredoxin (Trx1) in human lens epithelial cells under oxidative stress.

METHODS. The human TXN gene was cloned from a human lens cDNA library. Trx1 activity was measured by insulin reduction assay. For study of the upregulation of Trx1, 1.6 million human lens epithelial cells (HLE B3) were exposed to H2O2 (0.1 mM) for 0, 5, 10, 15, 20, and 30 minutes. The cells were lysed with lysis buffer and used for Trx1 activity assay, Western blot analysis, and real-time PCR.

RESULTS. The sequence of the human lens TXN gene was identical with that of other human tissues. Recombinant Trx1 was sensitive to iodoacetic acid but showed strong resistance to oxidation (0.1 mM H2O2) at its approximate physiological protein level. Western blot analysis and assay of Trx activity revealed that Trx1 was expressed in HLE B3 cells and localized in epithelial, cortical, and nuclear regions of human and porcine lenses. In vivo oxidative stress of HLE B3 cells resulted in a 35% upregulation of the level of Trx1 protein after 10 minutes of H2O2 treatment. Real-time PCR analysis showed an increase of approximately 50% in the level of Trx1 mRNA under the same oxidative stress conditions.

CONCLUSIONS. The upregulation of Trx1 in HLE B3 cells under oxidative stress and the presence of Trx1 in the lens suggest that the thioredoxin system may be an effective defense system that protects the lens against oxidative stress.




This article has been cited by other articles:


Home page
 IOVSHome page
S. Lofgren, M. R. Fernando, K.-Y. Xing, Y. Wang, C. A. Kuszynski, Y.-S. Ho, and M. F. Lou
Effect of Thioltransferase (Glutaredoxin) Deletion on Cellular Sensitivity to Oxidative Stress and Cell Proliferation in Lens Epithelial Cells of Thioltransferase Knockout Mouse
Invest. Ophthalmol. Vis. Sci., October 1, 2008; 49(10): 4497 - 4505.
[Abstract] [Full Text] [PDF]

Home page
 IOVSHome page
S. Moon, M. R. Fernando, and M. F. Lou
Induction of Thioltransferase and Thioredoxin/Thioredoxin Reductase Systems in Cultured Porcine Lenses under Oxidative Stress
Invest. Ophthalmol. Vis. Sci., October 1, 2005; 46(10): 3783 - 3789.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2003 by the Association for Research in Vision and Ophthalmology