Uber-responsive peptide-based sensors of signaling proteins.
Uber-responsive peptide-based sensors of signaling proteins.
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DOI：
10.1002/anie.200903717
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发表时间：
2009
影响因子：
16.6
通讯作者：
Lawrence, David S.
中科院分区：
文献类型：
--
作者：
Sharma, Vyas;Lawrence, David S.
一个错综复杂的细胞内生化途径网络是生命具有非凡适应性的原因。信号通路通过识别和响应环境变化，使细胞能够保持生命所需的细胞内条件。然而，这些途径也可以以对抗的能力发挥作用，字面上是在攻击寄主有机体。癌细胞在不适当的时间、不适当的地方分裂，并像它们的细菌同行一样，发展出巧妙的生化机制来击败治疗剂。20世纪的生物化学主要致力于蛋白质的分离和随后的体外表征。然而，更深入地了解蛋白质作用和细胞行为之间的关系，最好是通过研究感兴趣的蛋白质在其自然环境中获得的。细胞的生物化学，而不是试管的生物化学，驱动着面对环境挑战的适应性，无论是自然的还是人工的。我们对细胞生物化学的理解在很大程度上得益于荧光方面的非凡进步，特别是在技术和分子生物学领域。然而，第三个学科的最新进展，即合成，提供了对未来事物的形状的一瞥。绿色荧光蛋白(GFP)及其许多基因改变的结构已经使细胞生物学发生了革命性的变化。[1]将GFP添加到任何给定蛋白质的N或C末端是直接的，就像新构建的物种在活细胞中的表达一样。蛋白质会随着细胞分裂周期的变化而改变其位置吗？它的降解速度有多快，或者它在细胞内的扩散速度是多少？GFP也被用来检测附加蛋白质的动态化学变化。例如，含有FRET配对的GFP类似物(典型的青色荧光蛋白和黄色荧光蛋白；FRET=荧光共振能量转移)的结构已被设计为各种生物活性物质的传感器，如cAMP、[2]钙、[3]和蛋白激酶。[4]然而，尽管荧光蛋白的影响不能被夸大，GFP也不是没有局限性。GFP很大，因此可以改变它们所附加的蛋白质的生物行为(图1)。它们的大小也妨碍了荧光定位到所研究蛋白质上的特定亚区(例如，与活性部位相邻)。此外，除了蛋白质分解或某些其他形式的降解外，GFP通常对当地环境的变化没有反应。事实上，即使是双标签结构中的FRAT更改通常也不到50%。相比之下，小的荧光团不太可能干扰生物活性，它们提供了更大程度的
An intricate intracellular web of biochemical pathways is responsible for the remarkable adaptability of life. Signaling pathways enable cells to preserve the intracellular conditions required for life by recognizing and responding to environmental change. However, these pathways can also act in a confrontational capacity, literally turning on the host organism. Cancer cells divide at inappropriate times, in inappropriate places, and, like their bacterial counterparts, develop ingenious biochemical mechanisms to defeat therapeutic agents. Biochemistry in the 20th century was primarily devoted to the isolation of proteins and their subsequent characterization in vitro. However, a deeper understanding of the relationship between protein action and cellular behavior is best obtained by studying the protein of interest in its natural environment. The biochemistry of the cell, as opposed to the biochemistry of the test tube, drives adaptability in the face of environmental challenges, be they natural or artificial. Our understanding of cellular biochemistry has been greatly assisted by extraordinary advances in fluorescence, particularly in the areas of technology and molecular biology. However, recent progress from a third discipline, namely synthesis, offers a glimpse of the shape of things to come.Green fluorescent protein (GFP) and its many genetically altered constructs have revolutionized cell biology.[1] Appending GFPs to the N or C terminus of any given protein is straightforward, as is the expression of the newly constructed species in living cells. Does a protein change its location as a function of the cell division cycle? How rapidly is it degraded or what is its intracellular diffusion rate? GFPs have been used to detect dynamic chemical changes to the appended protein as well. For example, constructs containing FRET-paired GFP analogues (typically cyan fluorescent protein and yellow fluorescent protein; FRET= fluorescence resonance energy transfer) have been designed that serve as sensors for a variety of bioactive agents, such as cAMP,[2] Ca2+,[3] and protein kinases.[4] However, although the impact of fluorescent proteins cannot be overstated, GFPs are not without limitations. GFPs are large and thus can alter the biological behavior of the protein to which they are appended (Figure 1). Their large size also precludes localization of fluorescence to a specific subdomain (eg adjacent to the active site) on the protein under study. Furthermore, with the exception of proteolysis or some other form of degradation, GFPs are generally unresponsive to changes in their local environment. Indeed, even FRET changes in doubly labeled constructs are typically less than 50%. By contrast, small fluorophores are less likely to perturb biological activity, they provide a greater degree of
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影响因子：
14.8
作者：
Palmer, Amy E.;Tsien, Roger Y.
通讯作者：
Tsien, Roger Y.
影响因子：
15
作者：
Vázquez, ME;Nitz, M;Imperiali, B
通讯作者：
Imperiali, B
影响因子：
15
作者：
Sainlos M;Iskenderian WS;Imperiali B
通讯作者：
Imperiali B
DOI：
10.1016/j.bbapap.2007.07.016
发表时间：
2008-01-01
影响因子：
3.2
作者：
Shanna, Vyas;Wang, Qunzhao;Lawrence, David S.
通讯作者：
Lawrence, David S.
影响因子：
15
作者：
Loving G;Imperiali B
通讯作者：
Imperiali B