Design and synthesis of simplified energy-converting proteins.
Design and synthesis of simplified energy-converting proteins.
复制标题
简化的能量转换蛋白质的设计和合成。
DOI：
10.1042/bst0220689
复制
发表时间：
1994
影响因子：
3.9
通讯作者：
Dutton,PL
中科院分区：
文献类型：
--
作者：
Farid,RS;Robertson,DE;Moser,CC;Pilloud,D;DeGrado,WF;Dutton,PL
揭示生物化学结构和功能之间关系的传统方法是从分离相对纯净、稳定和有活性的蛋白质开始的。然后用光谱学和物理技术对这种制备进行检查，从而了解蛋白质的运作方式及其结构。如果蛋白质足够纯净，而且不是太大太复杂，那么通过使用蛋白质折叠模型，初级氨基酸序列可以提示一个整体结构。在某些情况下，可以培养出二维或三维的蛋白质晶体，这样衍射实验就可以缩小结构的可能性。关于结构如何转化为功能的假设可以通过从其他物种中纯化类似蛋白来进行有限的测试。如果这些基因能够被分离并转移到合适的生物体中，就可以制备大量的蛋白质，并且可以通过突变来操纵氨基酸序列。这些传统方法的缺点往往是显而易见的。许多具有最有趣功能的蛋白质在其复杂性方面是巨大而笨拙的：氧化磷酸化的电子传递蛋白很好地说明了这一点[11]。催化位点的位置可能被仅用于组装稳定结构的长序列所掩盖，或者仅仅是适应另一功能的进化残留物。在脊椎动物物种中，人们经常遇到大的蛋白质亚基，似乎只提供催化位点的微小调节。此外，这些复杂蛋白质的大小和物理性质往往阻碍了衍射研究的晶体生长。
The traditional approach to uncovering the relationship between biochemical structure and function begins with the isolation of a relatively pure, stable and active protein. This preparation is then examined with a battery of spectroscopic and physical techniques providing some idea of how the protein operates and what its structure might be. If the protein is pure enough and not too large and complex, the primary amino acid sequence can suggest, through use of protein folding models, an overall structure. In some cases, two-or three-dimensional protein crystals can be grown so that diffraction experiments can narrow down the structural possibilities. Ilypotheses about how the structure translates into function can be tested in a limited way by purifying analogues of the protein from other species. If the genes can be isolated and moved into a suitable organism, larger quantities of the protein can be prepared and manipulation of the amino acid sequence through mutation is possible. The shortcomings of these traditional methods are often painfully apparent. Many of the proteins with the most intriguing function are large and unwieldly in their complexity: the electrontransfer protein of oxidative phosphorylation illustrate this well [11. The location of catalytic sites may be obscured by long sequences that serve only to assemble a stable structure, or are simply evolutionary remnants once adapted to another function. In vertebrate species, one often encounters large protein subunits that seem to provide only minor regulation of the catalytic site. In addition, the size and physical properties of these complex proteins often preclude crystal growth for diffraction studies.