Fluorescent Proteins: Crystallization, Structural Determination, and Nonnatural Amino Acid Incorporation.
Fluorescent Proteins: Crystallization, Structural Determination, and Nonnatural Amino Acid Incorporation.
复制标题
荧光蛋白：结晶、结构测定和非天然氨基酸掺入。
DOI：
10.1007/978-1-0716-2667-2_5
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Ahmed RD
中科院分区：
文献类型：
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作者：
Ahmed RD
荧光蛋白通过作为基因编码标签的使用，彻底改变了细胞生物学和细胞成像。结构生物学在理解它们独特的荧光特性如何通过发色团的形成表现出来以及光谱特性如何通过相互作用网络进行调节方面至关重要。这些知识反过来又导致了具有新的和改进的特性的新变体的构建。在这里，我们描述的过程中，荧光蛋白的结构确定，从重组蛋白的生产开始的分子置换的结构确定。我们还描述了如何纳入和确定含有非天然氨基酸的蛋白质的结构。蛋白质工程的最新进展导致遗传密码的重新编程，以允许在设计的残基位置掺入新的化学物质，荧光蛋白处于该领域结构研究的最前沿。这种新化学对蛋白质结构的影响仍然有限;积累更多的蛋白质结构无疑将提高我们对具有新化学功能的蛋白质的理解和能力。
Fluorescent proteins have revolutionized cell biology and cell imaging through their use as genetically encoded tags. Structural biology has been pivotal in understanding how their unique fluorescent properties manifest through the formation of the chromophore and how the spectral properties are tuned through interaction networks. This knowledge has in turn led to the construction of novel variants with new and improved properties. Here we describe the process by which fluorescent protein structures are determined, starting from recombinant protein production to structure determination by molecular replacement. We also describe how to incorporate and determine the structures of proteins containing non-natural amino acids. Recent advances in protein engineering have led to reprogramming of the genetic code to allow incorporation of new chemistry at designed residue positions, with fluorescent proteins being at the forefront of structural studies in this area. The impact of such new chemistry on protein structure is still limited; the accumulation of more protein structures will undoubtedly improve our understanding and ability to engineer proteins with new chemical functionality.