BIOORGANIC STUDIES OF RETINAL PROTEINS

对视网膜蛋白质的研究涵盖了极其广泛的科学领域。 因为它完全是跨学科性质的；没有一个单一的学科可以 解决尚待解决的众多问题。的主要关注点 我们的研究是合成一些量身定做的视网膜类似物并使用 以澄清视网膜一般区域的具体方面 蛋白质参与其中。 (1)漂白适应：这是一个知之甚少的领域 而不是基于分子结构的视觉转导。这是因为 要处理的结构句柄要少得多，而且还因为 与视觉转导相比，视野远没有那么先进。然而， 从具有锁定的11顺-烯结构的模拟开始，我们有 找到了几条结构线索，这将使我们能够进一步设计 分子来阐明这一重要领域的机制。 (2)视觉传导的触发过程。在最后一次拨款期间 期间，我们研究了 转导过程。我们的下一个目标是探索光化学和 通过非常快速的光谱测量，非常早期的变化。 (3)主要努力是简化光亲和标记过程， 特别是以视紫红质为代表的膜蛋白。非常 很少有研究导致氨基酸的最终测序，因为 在分离多肽片段时遇到困难。我们会 继续简化分析方案。我们已经成功地 用视紫红质成功地表征了两种交联型氨基酸 (Rh)和细菌视紫红质(BR)。然而，这导致了进一步的 有趣的问题。在Rh的情况下，什么是绝对意义上的 发色团的扭曲，在BR的情况下，哪一个是正确的 结合部位内的取向或发色团。这些问题 将主要通过含有以下物质的类似物的固体核磁共振来解决 特定的标记，并应用一些非常规的核磁共振技术。 (4)我们实验室生产的大约100个模拟物中，大部分是用于 光谱学的目的。我们现在将按顺序测量酶的活性 为了进一步了解结构、电子和其他因素 与视觉传导有关。 (5)以视网膜模拟物为模板生产单抗 在人造体内携带适当分布电荷的抗体 11-顺式视网膜的结合部位；这提供了另一种方法 研究配体与其相互作用的静电效应 结合部位。

The studies on retinal proteins cover an extremely broad range of science because of its totally interdisciplinary nature; no single discipline can solve the numerous problems remaining to be answered. The main focus of our studies is to synthesize a number of tailored retinal analogs and use them to clarify specific aspects in the general area in which retinal proteins are involved. (1) Bleaching adaptation : This is a field which is much less understood than visual transduction on a molecular structural basis. This is because there are far fewer structural handles to deal with, and also because the field is far less advanced in comparison to visual transduction. However, starting with an analog which has a locked 11-cis-ene structure, we have found several structural clues which will allow us to design further molecules to clarify the mechanism of this important area. (2) The triggering process of visual transduction. During the last grant period, we studied the process and structural requirements of the transduction process. Our next aim is to explore the photochemistry and very early stage changes through very fast spectroscopic measurements. (3) A major effort is to streamline the process of photoaffinity labeling, particularly that of membrane proteins exemplified by the rhodopsins. Very few studies have led to the final sequencing of amino acids because of difficulties encountered in separation of the peptide fragments. We will continue to simplify the analytical protocols. We have managed to successfully characterize cross-linked amino acids both with rhodopsin (Rh) and bacteriorhodopsin (BR). However, this has led to further interesting questions. In the case of Rh, what is the absolute sense of twist of the chromophore, and in the case of BR, which is the correct orientation or the chromophore within the binding site. These problems will be addressed mainly through solid state NMR of analogs containing specific labels, and applying some non routine NMR techniques. (4) Most of the approximately 100 analogs made in our lab was for the purpose of spectroscopy. We will now measure enzymatic activities in order to gain further insight into the structural, electronic add other factors involved in visual transduction. (5) Retinal mimics will be used as templates to produce monoclonal antibodies that carry suitably distributed charges within artificial binding sites of 11-cis-retinal; this provides another approach for investigating electrostatic effects operating between the ligand and its binding site.