Journal
FEBS LETTERS
Volume 579, Issue 24, Pages 5414-5418Publisher
WILEY
DOI: 10.1016/j.febslet.2005.09.002
Keywords
11-cis retinal; isomerase; mutagenesis; retinal dystrophy; vitamin A; visual cycle
Funding
- NEI NIH HHS [EY012231, EY015650] Funding Source: Medline
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We have recently reported that RPE65 from the retinal pigment epithelium is the isomerohydrolase, a critical enzyme in the visual cycle for regeneration of 11-cis retinal, the chromophore for visual pigments. Here, we demonstrated that mutation of any one of the absolutely conserved four histidine and one glutamic acid residues to alanine in RPE65 abolished its isomerohydrolase activity. Substitution of the conserved glutamic acid with glutamine also resulted in loss of the activity. Moreover, these mutations significantly reduced protein stability of RPE65. These results indicate that these conserved residues are essential for the isomerohydrolase activity of RPE65 and its stability. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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