title: Identification of conserved histidines and glutamic acid as key residues for isomerohydrolase activity of RPE65, an enzyme of the visual cycle in the retinal pigment epithelium.
abstract: 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.
authors.affiliation: Department of Medicine Endocrinology, The University of Oklahoma Health Sciences Center, BSEB 328B, Oklahoma City, 73104, USA.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Gennadiy
authors.initials: G
authors.lastname: Moiseyev
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.forename: Jian-xing
authors.initials: JX
authors.lastname: Ma
mesh_terms: D002352:Carrier Proteins; D002384:Catalysis; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism*; D006801:Humans; D006867:Hydrolases / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010857:Pigment Epithelium of Eye / Q000502:physiology*; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D002352:Carrier Proteins; D005136:Eye Proteins; D018698:Glutamic Acid; D006639:Histidine; D006867:Hydrolases; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase