Vogel Reiner, Siebert Friedrich, Lüdeke Steffen, Hirshfeld Amiram, Sheves Mordechai
Biophysics Group, Institut für Molekulare Medizin und Zellforschung, Albert-Ludwigs-Universität Freiburg, Hermann-Herder-Strasse 9, D-79104 Freiburg, Germany.
Biochemistry. 2005 Sep 6;44(35):11684-99. doi: 10.1021/bi0508587.
Activation of the visual pigment rhodopsin is initiated by isomerization of its retinal chromophore to the all-trans geometry, which drives the conformation of the protein to the active state. We have examined by FTIR spectroscopy the impact of a series of modifications at the ring of retinal on the activation process and on molecular interactions within the binding pocket. Deletion of ring methyl groups at C1 and C5 or replacement of the ring in diethyl or ethyl-methyl acyclic analogues resulted in partial agonists, for which the conformational equilibrium between the Meta I and Meta II photoproduct is shifted from the active Meta II side to the inactive Meta I side. While the Meta II states of these artificial pigments had a conformation similar to those of native Meta II, the Meta I states were different. Modifications on the ring of retinal had a particular impact on the interaction of Glu 122 within the ring-binding pocket and are shown to interfere with the Glu 134-mediated proton uptake during formation of Meta II. We further found, upon partial deletion of ring constituents, a decrease of the entropy change of the transition from Meta I to Meta II by up to 50%, while the concomitant reduction of the enthalpy term was less pronounced. These findings underline the particular importance of the ring and the ring methyl groups and are discussed in a model of receptor activation.
视觉色素视紫红质的激活是由其视黄醛发色团异构化为全反式构象引发的,这驱使蛋白质的构象转变为活性状态。我们通过傅里叶变换红外光谱法研究了视黄醛环上一系列修饰对激活过程以及结合口袋内分子相互作用的影响。在C1和C5处缺失环甲基或在二乙基或乙基 - 甲基无环类似物中替换环会产生部分激动剂,对于这些部分激动剂,Meta I和Meta II光产物之间的构象平衡从活性的Meta II侧转移到非活性的Meta I侧。虽然这些人工色素的Meta II状态具有与天然Meta II相似的构象,但Meta I状态却不同。视黄醛环上的修饰对环结合口袋内Glu 122的相互作用有特别影响,并显示在Meta II形成过程中干扰Glu 134介导的质子摄取。我们进一步发现,在部分缺失环成分后,从Meta I到Meta II转变的熵变降低了高达50%,而焓项的相应减少则不太明显。这些发现强调了环和环甲基的特殊重要性,并在受体激活模型中进行了讨论。