Readily accessible bicyclononynes for bioorthogonal labeling and three-dimensional imaging of living cells.
Readily accessible bicyclononynes for bioorthogonal labeling and three-dimensional imaging of living cells.
复制标题
DOI：
10.1002/anie.201003761
复制
发表时间：
2010-12-03
影响因子：
16.6
通讯作者：
van Delft, Floris L.
中科院分区：
文献类型：
--
作者：
Dommerholt, Jan;Schmidt, Samuel;Temming, Rinske;Hendriks, Linda J. A.;Rutjes, Floris P. J. T.;van Hest, Jan C. M.;Lefeber, Dirk J.;Friedl, Peter;van Delft, Floris L.
在生物分子(蛋白质、脂类、多糖)的天然环境中成像和跟踪生物分子的化学生物学工具的出现为细胞过程提供了独特的见解，这是传统的生化或分子生物学工具无法实现的。[1]基于非生物功能标签和设计的化学探针之间的高度选择性反应，生物分子的生物正交标记已被证明对多糖[2]和脂类[3]的检测和研究特别有用。关于非生物标记，叠氮化合物因其直接的化学引入、小尺寸和相对惰性而被广泛使用。[4]叠氮化物在铜(I)存在的情况下与末端乙炔快速而清洁地反应的发现，在生命和材料科学中得到了巨大的应用。[5]然而，由于通常使用高达20mol%的铜(I)物种，这种点击化学不适合于在不损害细胞功能的情况下标记生命系统。[6]除此之外，铜的存在可能会导致寡核苷酸[7]和多糖[8]的降解。为了避免使用有毒金属，已经开发了几种无金属生物正交标记反应。尤其是，膦已被用于叠氮化物的共价连接，一种称为Staudinger连接的过程。[10]然而，由于膦的氧敏感性，最近的化学连接的焦点正转移到应变促进的与环辛炔的环加成反应(图式1a)。叠氮化物在所谓的应变促进的炔-叠氮环加成反应(SPAAC)中以高反应速率与环辛炔反应。[12]无金属生物正交反应的工具箱最近由我们的研究小组[13]和其他人[14]进一步扩展，证明了环辛炔与硝酮进行更快速的应变促进环加成反应(SPANC)，这一过程被发现适用于蛋白质的双重、不可逆和特定位置的N-末端修饰。[13]然而，无金属环加成反应在生命科学中的广泛应用，由于商业供应有限和制备最常见的环辛烷的合成路线较长而受到阻碍(方案1b)。例如，生成第二代Difo(1)需要8个合成步骤，DIBAC(3)、[16]需要9个步骤，Barac(4)、[17]需要7个步骤，而产率通常很低(2的产率为10%，4的产率为16%)。额外的修饰，如二苯并环化(化合物2-4)，增加了亲脂性，因此可能导致与蛋白质的非特异性结合。作为一种新型的环紧链炔，用于与叠氮化物和硝酮的无金属环加成反应。通过1，5-环辛二烯的环丙烷化反应，以非常简单的方法得到了具有Cs对称性的双环酮衍生物，并且在应变促进的环加成反应中表现出了良好的反应动力学。BCN的功能化衍生物被应用于蛋白质和多糖的标记，以及活体黑色素瘤细胞的三维可视化。
The advent of chemical biology tools for imaging and tracking of biomolecules (proteins, lipids, glycans) in their native environment is providing unique insights into cellular processes that are not achievable with traditional biochemical or molecular biology tools.[1] Bioorthogonal labeling of biomolecules has proven particularly useful for the detection and study of glycans [2] and lipids,[3] based on a highly selective reaction between an abiotic functional tag and a designed chemical probe. With respect to the abiotic tag, azide has been used extensively because of its straightforward chemical introduction, small size, and relative inertness.[4] The finding that azides react rapidly and cleanly with terminal acetylenes in the presence of copper (I), the quintessential “click” reaction, has found tremendous application in life and material sciences.[5] However, because up to 20mol% of copper (I) species is typically used, such click chemistry is not suitable for labeling of living systems without compromising cell function.[6] Apart from that, the presence of copper may induce oligonucleotide [7] and polysaccharide [8] degradation. To avoid the use of toxic metals, several metal-free bioorthogonal labeling reaction have been developed.[9] In particular, phosphines have been used for covalent ligation to azides, a procedure known as Staudinger ligation.[10] However, owing to the oxygen sensitivity of phosphines, recent focus of chemical ligation is shifting towards strain-promoted cycloaddition reactions with cyclooctynes (Scheme1a).[11] Most prominently, azides were find to react with cyclooctynes with high reaction rates in a so-called strain-promoted alkyne–azide cycloaddition (SPAAC).[12] The toolbox of metal-free bioorthogonal reactions was most recently further expanded by our research group [13] and others,[14] by demonstrating that cyclooctynes undergo even more rapid strainpromoted cycloaddition with nitrones (SPANC), a procedure that was found suitable for dual, irreversible, and site-specific N-terminal modification of proteins.[13] The broad application of metal-free cycloaddition in life sciences is, however, hampered by the limited commercial availability and lengthy synthetic routes for preparation of the most common cyclooctynes (Scheme 1b). For example, eight synthetic steps are required to generate second-generation DIFO (1),[15] nine steps for DIBAC (3),[16] and seven steps for BARAC (4),[17] while yields are usually low (10% for 2,[18] 16% for 4). Additional modifications, such as dibenzoannulation (compounds 2–4), increase lipophilicity and may, therefore lead to non-specific binding to proteins.[17] Here we report bicyclo [6.1. 0] nonyne (BCN) as a novel ring-strained alkyne for metal-free cycloaddition reactions with azides and nitrones. Bicyclononyne derivatives, which were obtained in a highly straightforward process through cyclopropanation of 1, 5-cyclooctadiene, are Cs symmetrical and display excellent reaction kinetics in strain-promoted cycloaddition reactions. Functionalized derivatives of BCN were applied in the labeling of proteins and glycans, as well as in the three-dimensional visualization of living melanoma cells.
登录
查看更多内容
影响因子：
4.8
作者：
Link, AJ;Tirrell, DA
通讯作者：
Tirrell, DA
影响因子：
5.6
作者：
Chiang, Chi-Hsiang;Wang, Chie-Hong;Hung, Wen-Chun
通讯作者：
Hung, Wen-Chun
影响因子：
16.6
作者：
Lutz, Jean-Francois
通讯作者：
Lutz, Jean-Francois
影响因子：
15
作者：
Lallana, Enrique;Fernandez-Megia, Eduardo;Riguera, Ricardo
通讯作者：
Riguera, Ricardo
影响因子：
15
作者：
Agard, NJ;Prescher, JA;Bertozzi, CR
通讯作者：
Bertozzi, CR