Aromatic retinal analogues and their interaction with cattle opsin.
Aromatic retinal analogues and their interaction with cattle opsin.
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芳香族视黄醛类似物及其与牛视蛋白的相互作用。
DOI：
10.1021/bi00561a008
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发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
Liu,RS
中科院分区：
文献类型：
--
作者：
Matsumoto,H;Asato,AE;Denny,M;Baretz,B;Yen,YP;Tong,D;Liu,RS
材料和方法所有芳香族视黄酸均按照C15 + C5路线合成（Bharucha &韦登，1953）。以下是一般程序的简要说明。以适当取代的苯甲醛与4-（二乙基膦酰基）-3-甲基巴豆腈的锂衍生物（E/Z= 1）在H_4呋喃2-HMPA-己烷中反应，以良好的产率（60-85%）制备5-芳基-3-甲基-2，4-丁烯腈。在苯-己烷中用二异丁基氢化铝（DIBAH）还原5-芳基二烯腈（1.5小时，0 ℃至室温），然后进行湿硅胶处理（Kini等人，1979），以中等产率（50-65%）得到相应的5-芳基-3-甲基-2，4-双烯醛。随后重复上述二烯醛与膦酸酯（EtO）2 POCHC（CH 3）= CHCN的扩链反应，然后DIBAH还原成所需的芳族视黄醛类似物VI和VII，以相当的总产率进行醛V是由（2 E，4 E）-3-甲基-5-（4-甲氧基苯基）-2-甲基-5-（4-甲氧基苯基）-2-（三氟甲基）苯缩合制得的。将（2，4，6-三甲基苯基）-2，4-双烯醛与（E）-3-甲基-4-（二乙基膦酰基）巴豆酸甲酯的锂衍生物反应，然后（a）氢化铝锂还原和（B）二氧化锰氧化并通过从乙腈中重结晶纯化。醛VIII是通过对氰基-甲苯基膦酸酯与β-紫罗兰酮的均三甲苯类似物缩合，随后用DIBAH部分还原而制备的。用于合成IX的起始材料是α-甲基肉桂醛。牛视蛋白及其与芳香族视网膜的相互作用通过常规方法纯化视杆外节（ROS）（Hubbard等人，1971;松本等人，1978）从购自Hormel的冷冻牛视网膜获得。纯化的ROS用蒸馏水洗涤两次并冻干。然后将它们用冷石油醚洗涤数次。用溶于10 mM 7V-2-羟乙基哌嗪-N，N-2-乙磺酸（Hepes）缓冲液（pH 7.0）中的1%毛地黄皂苷从所得ROS中提取视蛋白。
Materials and MethodsAll aromatic retináis were synthesized according to the C] 5+ C5 route (Bharucha & Weedon, 1953). Thefollowing is a brief description of the general procedure. The preparation of 5-aryl-3-methyl-2, 4-pentadienonitriles was effected in good yields (60-85%) by the reaction of appropriately substituted benzaldehydes with the lithio derivative of 4-(diethyl-phosphono)-3-methylcrotononitrile (£/Z= 1) in H4furan2-HMPA-hexane. Reduction of the 5-aryldienonitriles with diisobutylaluminum hydride (DIBAH) in benzene-hexane (1.5 h, 0 C to room temperature), followed by a wet silica gel workup (Kini et al., 1979), afforded the corresponding 5-aryl-3-methyl-2, 4-pentadienals in moderate yields (50-65%). Subsequent repetition of the chain extension reaction of the above dienals with the phosphonate (EtO) 2POCHC (CH3)= CHCN, followed by DIBAH reduction to the desired aromatic retinal analogues VI and VII, was effected in fair overall yields (after recrystallization from benzene-hexane or ethyl ace-tate-hexane).Aldehyde V was prepapred by the condensation of (2£, 4£)-3-methyl-5-(2, 4, 6-trimethylphenyl)-2, 4-pentadienal with the lithium derivative of methyl (£)-3-methyl-4-(di-ethylphosphono) crotonate, followed by (a) lithium aluminum hydride reduction and (b) manganese dioxideoxidation and purification by recrystallization from acetonitrile. Aldehyde VIII was prepared by condensation of p-cyano-tolylphosphonate with the mesityl analogue of/3-ionone, fol-lowed by partial reduction with DIBAH. The starting material for the synthesis of IX was a-methylcinnamaldehyde. Cattle Opsin and Its Interaction with Aromatic Retináis. Rod outer segments (ROS) were purified by a conventional method (Hubbard et al., 1971; Matsumoto et al., 1978) from frozen cattle retinas purchased from Hormel. The purified ROS were washed twicewith distilled water and lyophilized. Then they were washed with chilled petroleum ether several times. Opsin was extracted from the resultant ROS with 1% digitonin dissolved in 10 mM 7V-2-hydroxyethylpiperazine-A^-2-ethanesulfonic acid (Hepes) buffer, pH 7.0.