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Identification of DES1 as a vitamin A isomerase in Müller glial cells of the retina

Nature Chemical Biology volume 9, pages 30–36 (2013)Cite this article

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Abstract

Absorption of a light particle by an opsin-pigment causes photoisomerization of its retinaldehyde chromophore. Restoration of light sensitivity to the resulting apo-opsin requires chemical re-isomerization of the photobleached chromophore. This is carried out by a multistep enzyme pathway called the visual cycle. Accumulating evidence suggests the existence of an alternative visual cycle for regenerating opsins in daylight. Here we identified dihydroceramide desaturase-1 (DES1) as a retinol isomerase and an excellent candidate for isomerase-2 in this alternative pathway. DES1 is expressed in retinal Müller cells, where it coimmunoprecipitates with cellular retinaldehyde binding protein (CRALBP). Adenoviral gene therapy with DES1 partially rescued the biochemical and physiological phenotypes in Rpe65−/− mice lacking isomerohydrolase (isomerase-1). Knockdown of DES1 expression by RNA interference concordantly reduced isomerase-2 activity in cultured Müller cells. Purified DES1 had very high isomerase-2 activity in the presence of appropriate cofactors, suggesting that DES1 by itself is sufficient for isomerase activity.

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Figure 1: Immunohistochemical and immunoblotting analysis showing DES1 expression in the retina and RPE.
Figure 2: Isomerase-2 activities of human membrane desaturases.
Figure 3: Levels of visual chromophore and ERG amplitudes in Rpe65−/− mice after intravitreal injection of ad-DES1 or ad-RFP.
Figure 4: Coimmunoprecipitation of CRALBP with DES1 and effects of CRALBP on isomerase-2 activity.
Figure 5: Isomerase-2 activity of purified GST-DES1.
Figure 6: Proposed alternative visual cycle in Müller cells.

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Acknowledgements

We thank R. Radu for insightful suggestions throughout this project and valuable comments on the manuscript, J. Hu and D. Bok for assistance with immunocytochemistry, M. Redmond (National Eye Institute) for providing Rpe65−/− mice, and R. Crouch (Medical University of South Carolina) for the gift of 11-cis-RAL. This work was supported by a grant to G.H.T. from the US National Eye Institute (R01-EY11713). G.H.T. is the Charles Kenneth Feldman Professor of Ophthalmology.

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Authors and Affiliations

  1. Jules Stein Eye Institute, University of California, Los Angeles School of Medicine, Los Angeles, California, USA

    Joanna J Kaylor, Quan Yuan, Jeremy Cook, Shanta Sarfare, Jacob Makshanoff, Anh Miu, Anita Kim, Paul Kim, Samer Habib, C Nathaniel Roybal, Tongzhou Xu, Steven Nusinowitz & Gabriel H Travis

  2. Department of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California, USA

    Gabriel H Travis

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Contributions

G.H.T. conceived the project. G.H.T., J.J.K., C.N.R., Q.Y. and S.N. designed the experiments and interpreted the data. J.J.K., Q.Y., J.C., S.S., J.M., A.M., A.K., P.K., S.H., T.X. and C.N.R. performed the experiments. G.H.T. wrote the manuscript.

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Correspondence to Gabriel H Travis.

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Kaylor, J., Yuan, Q., Cook, J. et al. Identification of DES1 as a vitamin A isomerase in Müller glial cells of the retina. Nat Chem Biol 9, 30–36 (2013). https://doi.org/10.1038/nchembio.1114

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