*** binding properties of the C-terminal segment of retinol dehydrogenase 8

作     者：AndréHädicke Ana I.A.Coutinho François Otis Mustapha Lhor Line Cantin Manuel Prieto Normand Voyer Christian Salesse 

作者机构：PROTEO and CUO-RechercheCentre de recherche du CHU de Québec-UniversitéLavalHôpital St-SacrementQuébecCanada Centro Química-Física MolecularInstituto Superior TécnicoUniversidade de LisboaLisboaPortugal PROTEO and Dépt.ChimieFacultéSciences et génieUniversitéLavalQuébecCanada 

出 版 物：《Annals of Eye Science》 (眼科学年鉴（英文）)

年 卷 期：2018年第1期

页      面：428-428页

学科分类：081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 070303[理学-有机化学] 0703[理学-化学] 

主　　题：Peptide-membrane binding electrostatic interactions hydrophobic interactions fluorescence spectroscopy 

摘      要：Background:Retinol dehydrogenase 8(RDH8)is a 312-amino acid(aa)protein involved in the visual *** to the outer segment disk membranes of photoreceptors,it reduces all-trans-retinal to all-trans-retinol1 as one of the rate-limiting steps of the visual ***8 is a member of the short-chain dehydrogenase/reductase *** C-terminal segment allows its membrane-anchoring through the postulated presence of an amphipathicα-helix and of 1 to 3 acyl groups at positions 299,302 and *** secondary structure and membrane binding characteristics of RDH8 and its C-terminal segment have not yet been ***:To evaluate the membrane binding of RDH8,the full-length protein(aa 1-312),a truncated form(aa 1-296),its C-terminal segment(aa 281-312 and 297-312)as well as different additional variants of this segment were *** truncated protein binds membranes less efficiently than the full-length ***,the C-terminal segment of RDH8 is essential for the binding and has thus been further *** intrinsic fluorescence of tryptophan residues at positions 289 and 310 of the wild-type C-terminal segment of RDH8 and the mutants W289F,W310F and W310R have thus been used to determine their extent of binding to lipid vesicles and to monitor their local *** lipid vesicles composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine(POPC)or a mixture of POPC and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine(POPS)were used to mimic the phospholipid content of the outer segment disk membranes of ***:An increase in fluorescence intensity and in fluorescence lifetime is observed upon increasing the concentration of lipid *** data allowed calculating values of partition coefficient of the C-terminal segment of RDH8 varying between Kp=1.1 E6 to 1.7 *** is noteworthy that the observation of a more intense shift to lower wavelengths upon membrane binding of the mutant W310R and W310F indicates a deeper inco