BIOORGANIC STUDIES OF RETINAL PROTEINS

视网膜蛋白质的研究涉及的科学领域非常广泛 由于其完全跨学科的性质;没有一个学科可以 解决了许多有待解决的问题。的主要重点 我们的研究是合成一些定制的视网膜类似物， 他们澄清在视网膜的一般领域的具体方面， 蛋白质参与其中。 (1)漂白适应：这是一个领域，这是少得多的理解 比视觉传导更重要。这是因为 要处理的结构句柄要少得多，而且还因为 场远不如视觉转导先进。然而，在这方面， 从具有锁定的11-顺式-烯结构的类似物开始，我们得到 发现了几个结构线索，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。在上一次赠款期间 期间，我们研究了工艺和结构要求， 转导过程我们的下一个目标是探索光化学， 通过非常快速的光谱测量， (3)一个主要的努力是简化光亲和标记的过程， 特别是以视紫红质为例的膜蛋白的活性。非常 很少有研究导致氨基酸的最终测序，因为 在分离肽片段时遇到的困难。我们将 继续简化分析方案。我们已经设法 成功地表征了与视紫红质交联氨基酸 (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， 发色团的扭曲，在BR的情况下，这是正确的 方向或结合位点内的发色团。这些问题 将主要通过固态NMR的类似物， 特异性标记，并应用一些非常规的核磁共振技术。 (4)在我们实验室制造的大约100种类似物中， 光谱学的目的我们现在将测量酶活性， 为了进一步了解结构，电子添加其他因素， 参与视觉传导。 (5)视网膜模拟物将被用作产生单克隆抗体的模板。 抗体携带适当分布的电荷， 结合位点的11-顺式-视网膜;这提供了另一种方法， 研究配体与其配体之间的静电效应， 结合位点

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.