Specificity of binding of all-trans-retinyl ester to RPE65.
Membrane-bound RPE65 (mRPE65) is a binding protein for all-trans-retinyl esters, which are the substrates for the isomerization reaction that completes the visual cycle. RPE65 is essential for rhodopsin regeneration and, hence, for vision. As RPE65 appears to be part of the rate-limiting pathway in the visual cycle, specific antagonists of the molecule will be important in evaluating its full physiological role. The protein is known to stereoselectively bind all-trans-retinyl esters (tREs), with dissociation constants in the 50 nM range. This study explores the overall binding specificity of RPE65 with respect to both retinoids and other isoprenoids in an effort to define the specificity of binding, and to begin the process of designing specific antagonists for it. The nature of the specificity directed toward the three main structural elements (retinoid, linker, and acyl moieties) in the tRE molecule is reported. In the all-trans-retinyl ester series, binding affinity increased as a function of the hydrophobicity of the fatty acyl group. In the linker region, binding affinities were little affected by amide, ketone, and ether replacements for the carboxy ester moiety of the naturally occurring tRE ligand. Finally, modifications in the all-trans-retinoid moiety are also tolerated. For example, E,E-farnesyl palmitate binds with approximately the same affinity as does all-trans-retinyl palmitate. Other isoprenoid analogues also bind, as do truncated retinoids in the beta-ionone series. Therefore, mRPE65 is a moderately specific retinoid binding protein directed at long chain all-trans-retinyl esters.
Maiti
Pranab
P
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 45 Shattuck Street, Boston, Massachusetts 02115, USA.
Gollapalli
Deviprasad
D
Rando
Robert R
RR
D000818:Animals; D002417:Cattle; D004952:Esters / Q000737:chemistry / Q000378:metabolism; D005136:Eye Proteins / Q000737:chemistry / Q000378:metabolism*; D015394:Molecular Structure; D010857:Pigment Epithelium of Eye / Q000737:chemistry; D011485:Protein Binding; D012176:Retinoids / Q000737:chemistry / Q000378:metabolism; D012177:Retinol-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D012243:Rhodopsin / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D014785:Vision, Ocular
D004952:Esters; D005136:Eye Proteins; D012176:Retinoids; D012177:Retinol-Binding Proteins; D012243:Rhodopsin