共查询到20条相似文献,搜索用时 156 毫秒
1.
2.
制备了两种不同官能团修饰的偶联亲和素的包裹钌联吡啶(RuBpy)二氧化硅荧光纳米探针A和探针B,并分别用于肝癌细胞的识别。通过反相微乳液法制备得到表面修饰不同官能团的纳米颗粒,然后通过亲和素与羧基化包裹RuBpy二氧化硅纳米颗粒相互连接而制备得到探针A;通过亲和素与PEG修饰的荧光二氧化硅纳米颗粒相互作用而制备得到探针B。与探针A不同的是,探针B通过一个长链PEG分子将亲和素与荧光二氧化硅纳米颗粒偶联在一起。利用免疫荧光成像法将这两种探针分别用于人肝癌细胞的识别,结果表明,含有长链PEG分子的探针B更能够有效地识别肝癌细胞表面肿瘤标志物癌胚抗原(CEA)。 相似文献
3.
本工作将罗丹明B分子通过共价结合的方式成功地包裹在二氧化硅纳米粒子中,制备的纳米粒子荧光强度和罗丹明B分子相比提高了1000倍.对此硅纳米荧光粒子进一步进行了链亲和素修饰,成功制备了可特异性结合生物素修饰蛋白的纳米荧光检测探针.以反相蛋白质芯片检测为模式,研究了此探针对微量蛋白的检测性能.实验中将不同微量浓度的人IgG固定于醛基修饰玻璃片表面,并加入生物素标记的抗人IgG,结果显示在800fg~100pg含量的微量蛋白检测中此纳米荧光探针具有良好的线性关系,最小蛋白检测量可达100fg.与商品化亲和素偶联cy3荧光探针对比分析发现,本方法制备的荧光探针对蛋白的检测灵敏度可提高8倍,且具有成本低,生物修饰简单等优点. 相似文献
4.
生物素修饰纳米银探针的制备及在蛋白芯片可视化检测中的应用 总被引:1,自引:0,他引:1
采用寡核苷酸为连接分子成功制备了生物素修饰的纳米银探针, 并建立了纳米银催化同种金属离子的特异性还原显色反应. 实验采用蛋白质芯片为分析工具, 以微量人IgG为蛋白分析模式研究了纳米银探针/氢醌/硝酸银体系的显色分析性能. 实验结果表明, 上述检测体系可对160 fg~100 pg含量范围内的微量蛋白显示可视化结果, 蛋白点的灰度值与其浓度具有良好的相关性, 最小蛋白检测量可达160 fg. 同时还开展了与商品化链亲和素纳米金/银增强试剂显色方法的对比实验, 结果表明, 本法制备的探针对蛋白的检出限降低了约40倍, 且具有存储稳定、反应快速等优点. 相似文献
5.
本研究基于原位水热合成和气相浸渍的方法分别合成了含有Br?nsted/Lewis酸的Beta分子筛和只含有Lewis酸的AlCl3@Si-Beta样品,并通过原位漫反射红外技术对比研究了Br?nsted酸和Lewis酸催化直链和侧链戊烯同分异构体转化作用机理的差异。结果表明,同时含有Br?nsted/Lewis酸的Beta分子筛中,Br?nsted酸起主要的活化作用,催化戊烯进行异构和叠合反应时均遵循经典的碳正离子机理;而AlCl3@Si-Beta中的Lewis酸不含氢质子或羟基,催化α-戊烯进行双键迁移和2-戊烯的顺反异构反应时遵循AB-AD机理,以类烯丙基物种作为中间体,无法催化戊烯进行骨架异构和叠合反应。 相似文献
6.
7.
基于分子发夹/荧光微球探针构建了一种侧向层析定量检测黄曲霉毒素B1(AFB1)的新方法。一条含有AFB1适配体的分子发夹与荧光微球偶联后,形成的标记探针被喷涂在试纸条的结合垫上。一条5'端含有链霉亲和素的寡核苷酸序列包被在硝酸纤维素膜的检测线上,另一条包含有AFB1适配体互补链的寡核苷酸序列包被在硝酸纤维素膜的质控线上。当待测样本中含有AFB1时,AFB1先与标记物结合垫处标记探针中的AFB1适配体结合,同时,标记探针中的DNA分子发夹‘茎’的双链被打开,AFB1与标记探针形成的复合物层析到反应膜的检测区时,被检测区的寡核苷酸序列捕获,检测区出现亮线。利用该原理,通过一个"off-on"的光信号,实现了对AFB1的高灵敏检测。实验结果表明,AFB1在0.1~50μg/L质量浓度范围内,检测线处的荧光强度(T)和质控线荧光强度(C)的比值与AFB1质量浓度呈良好的线性关系,检出限(S/N=3)达0.05μg/L,且具有很高特异性,可实现对实际样品中AFB1的准确检测。 相似文献
8.
通过物理吸附和共价作用机制, 制备两种链亲和素-磁性微粒, 即链亲和素-金磁微粒和链亲和素-氨基磁粒, 并对其在不同缓冲液中的稳定性进行研究; 采用酶抑制法测定两种链亲和素-磁性微粒对游离生物素的结合能力; 分别以紫外吸收和固相核酸杂交方法, 测定两种链亲和素-磁性微粒对生物素标记寡核苷酸探针的固定化容量及活性, 并与Dynabeads®M-270 Streptavidin进行比较. 结果表明: 通过物理吸附作用制备的链亲和素-金磁微粒, 适用于核酸杂交与检测常用的STE (Tris-NaCl-EDTA) 缓冲系统, 通过共价作用形成的链亲和素-氨基磁粒, 适用于STE和磷酸盐(PBS)缓冲系统; 1 mg链亲和素-金磁微粒和链亲和素-氨基磁粒对游离生物素的最大结合容量分别为4950和5115 pmol; 对生物素标记寡核苷酸探针(24 mer) 的结合容量分别为2839和2978 pmol, 测定结果均是Dynabeads®M-270 Streptavidin的6~7倍; 与FITC-标记互补寡核苷酸的杂交结果表明, 固定于链亲和素-磁性微粒表面的寡核苷酸探针保持了较好的生物学活性. 相似文献
9.
功能化纳米金增强的DNA电化学检测和序列分析 总被引:6,自引:0,他引:6
用冠以大量二茂铁的纳米金微粒 /抗生蛋白链菌素结合物为标记物 ,将其标记于生物素修饰的寡聚核苷酸片段上 ,制成了具有电化学活性和纳米金放大作用的DNA电化学生物传感器 .首先采用巯基DNA和巯基烷烃混合自组装膜制备了金修饰电极 ,将探针DNA分子固定在了电极表面 ,运用杂交原则结合靶点分子在电极表面形成了双螺旋的DNA链 ,然后借助抗生蛋白链菌素和生物素之间的强亲和作用 ,引入了功能化的纳米金 .通过伏安法测定了修饰在纳米金上的二茂铁的氧化还原电流 ,可以识别和测定溶液中互补的靶点DNA ,17 mer靶点DNA的浓度在 0 .0 0 1~ 10nmol/L范围内有线性关系 ,检测限可达 0 .75× 10 -12 mol/L . 相似文献
10.
柠檬醛及中间体的合成 总被引:1,自引:0,他引:1
以异戊烯醛和异戊烯醇为原料,经异戊烯酸催化缩醛化反应得到3-甲基-2-丁烯醛二异戊烯基缩醛,再经磷酸二氢铵催化消除反应得到顺/反-异戊二烯基-3-甲基丁二烯醚,并进一步热重排获得柠檬醛。研究了缩醛化反应条件和消除反应条件对转化率的影响。结果表明,以0.3%异戊烯酸为酸性催化剂,70~75℃共沸脱水反应8 h,异戊烯醛的单程转化率达到63%~64%,处理后可得含量为97.5%的3-甲基-2-丁烯醛二异戊烯基缩醛,收率96.8%;以0.2%~0.5%磷酸二氢铵为催化剂,100~130℃,2.66 kPa下反应并及时将反应产物蒸出,处理后得到含量为95.9%的顺/反-异戊二烯基-3-甲基丁二烯醚,收率97.0%。顺/反-异戊二烯基-3-甲基丁二烯醚在120~130℃重排反应1 h,其反应产物柠檬醛含量97.5%,收率90.2%。 相似文献
11.
In order to characterize binding sites of insecticidal compounds on GABA gated chloride channel,new photoaf-finity probe candidates based on 5e-t-butyl-2e-[4-(substituted-propynyl)phenyl]-1,3-dithiane for the noncompetitiveblocker(NCB)site of the γ-aminobutyric acid(GABA)-gated chloride channel were designed and synthesized,andtheir potency as an inhibitor on NCB was measured by 4'-ethynyl-4-n-[2,3-~3H_2]-propylbicycloorthobenzoate(~3HEBOB)assay.The synthesized compounds showed high inhibition activities with half maximum inhibition concen-trations(IC_(50))of lower than 35 nmol/L and were very stable in binding conditions as well photoreacted quickly at300 nm light.These new compounds are expected to be good photoaffinity labeling probes if radioisotope iodine isincorporated. 相似文献
12.
Dr. David J. Fallon Stephanie Lehmann Dr. Chun-wa Chung Alex Phillipou Dr. Christian Eberl Ken G. M. Fantom Dr. Francesca Zappacosta Dr. Vipulkumar K. Patel Dr. Marcus Bantscheff Prof. Christopher J. Schofield Prof. Nicholas C. O. Tomkinson Dr. Jacob T. Bush 《Chemistry (Weinheim an der Bergstrasse, Germany)》2021,27(71):17880-17888
We present a one-step Ugi reaction protocol for the expedient synthesis of photoaffinity probes for live-cell MS-based proteomics. The reaction couples an amine affinity function with commonly used photoreactive groups, and a variety of handle functionalities. Using this technology, a series of pan-BET (BET: bromodomain and extra-terminal domain) selective bromodomain photoaffinity probes were obtained by parallel synthesis. Studies on the effects of photoreactive group, linker length and irradiation wavelength on photocrosslinking efficiency provide valuable insights into photoaffinity probe design. Optimal probes were progressed to MS-based proteomics to capture the BET family of proteins from live cells and reveal their potential on- and off-target profiles. 相似文献
13.
《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2017,129(27):8043-8046
An approach to biogenetically overlooked areas of the isoprenoid chemical space is presented. This strategy is based on the generation of a cationic center in functionalized polyolefins by Lewis acid activation of a carbonyl group, rather than by electrophilic attack at a double bond. Starting from the monocyclic humulane trienone zerumbone, polycyclic sesquiterpenoid skeletons which are either not reported as natural products or biogenetically enigmatic in terms of the isoprenoid rule, were obtained by modulating the Lewis acid catalyst. In the course of these studies, the surprising formation of a strained E ‐cyclooctene motif was observed in a cyclization reaction. 相似文献
14.
The N‐acylsulfonamide group, known as a safety‐catch linker, has been applied to photoaffinity labeling (PAL) using a cinnamate‐type photocrosslinker to improve the efficiency of PAL‐based target identification. A bioorthogonal sulfo‐click reaction was used to stably link a photocrosslinker unit with N‐acylsulfonamide linkage to produce a photoactivatable probe without any protection. In addition, the crosslinked protein was selectively isolated with a small cinnamate tag via linkage disruption upon N‐alkylation. Furthermore, the tag moiety was photochemically converted to a stable coumarin derivative by losing a water molecule, which is a useful property in MS‐based identification. 相似文献
15.
Unravelling RNA–Substrate Interactions in a Ribozyme‐Catalysed Reaction Using Fluorescent Turn‐On Probes
下载免费PDF全文
![点击此处可从《Chemistry (Weinheim an der Bergstrasse, Germany)》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Ece Cazibe Gaffarogullari Dr. Peter Greulich Dr. Andrei Yu. Kobitski Dr. Alexander Nierth Prof. Dr. G. Ulrich Nienhaus Prof. Dr. Andres Jäschke 《Chemistry (Weinheim an der Bergstrasse, Germany)》2015,21(15):5864-5871
The Diels–Alder reaction is one of the most important C?C bond‐forming reactions in organic chemistry, and much effort has been devoted to controlling its enantio‐ and diastereoselectivity. The Diels–Alderase ribozyme (DAse) catalyses the reaction between anthracene dienes and maleimide dienophiles with multiple‐turnover, stereoselectivity, and up to 1100‐fold rate acceleration. Here, a new generation of anthracene‐BODIPY‐based fluorescent probes was developed to monitor catalysis by the DAse. The brightness of these probes increases up to 93‐fold upon reaction with N‐pentylmaleimide (NPM), making these useful tools for investigating the stereochemistry of the ribozyme‐catalysed reaction. With these probes, we observed that the DAse catalyses the reaction with >91 % de and >99 % ee. The stereochemistry of the major product was determined unambiguously by rotating‐frame nuclear Overhauser NMR spectroscopy (ROESY‐NMR) and is in agreement with crystallographic structure information. The pronounced fluorescence change of the probes furthermore allowed a complete kinetic analysis, which revealed an ordered bi uni type reaction mechanism, with the dienophile binding first. 相似文献
16.
Alyson M. Curry Elizabeth Barton Wenjia Kang Daniel V. Mongeluzi Yana Cen 《Molecules (Basel, Switzerland)》2021,26(1)
NAD+ (nicotinamide adenine dinucleotide)-dependent protein deacylases, namely, the sirtuins, are important cell adaptor proteins that alter cell physiology in response to low calorie conditions. They are thought to mediate the beneficial effects of calorie restriction to extend longevity and improve health profiles. Novel chemical probes are highly desired for a better understanding of sirtuin’s roles in various biological processes. We developed a group of remarkably simple activity-based chemical probes for the investigation of active sirtuin content in complex native proteomes. These probes harbor a thioacyllysine warhead, a diazirine photoaffinity tag, as well as a terminal alkyne bioorthogonal functional group. Compared to their benzophenone-containing counterparts, these new probes demonstrated improved labeling efficiency and sensitivity, shortened irradiation time, and reduced background signal. They were applied to the labeling of individual recombinant proteins, protein mixtures, and whole cell lysate. These cell permeable small molecule probes also enabled the cellular imaging of sirtuin activity change. Taken together, our study provides new chemical biology tools and future drug discovery strategies for perturbing the activity of different sirtuin isoforms. 相似文献
17.
《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2017,129(10):2788-2792
Biochemical studies of cellular processes involving polyubiquitin have gained increasing attention. More tools are needed to identify ubiquitin (Ub)‐binding proteins. We report diazirine‐based photoaffinity probes that can capture Ub‐binding proteins in cell lysates, and show that diazirines are preferable to aryl azides as the photo‐crosslinking group, since they decrease non‐selective capture. Photoaffinity probes containing at least two Ub units were required to effectively capture Ub‐binding proteins. Different capture selectivity was observed for probes containing diubiquitin moieties with different types of linkages, thus indicating the potential to develop linkage‐dependent probes for selectively profiling Ub‐binding proteins under various cellular conditions. 相似文献
18.
Novel biotin-tagged photoaffinity probes based on a trifunctional tertiary amine scaffold were synthesized and evaluated as vascular endothelial growth factor receptor-2 (VEGFR-2) inhibitors. Probes 3a-c inhibit VEGF induced proliferation in HUVE cells, with IC50 values of 29.7, 33.3, and 37.7 μM, respectively. Moreover, we identified the interaction of 3b with VEGFR-2 in photoaffinity labeling experiment using HUVE cells. 相似文献
19.
Qingyun Zheng Tian Wang Guo‐Chao Chu Chong Zuo Rui Zhao Xin Sui Linzhi Ye Yuanyuan Yu Jingnan Chen Xiangwei Wu Wenhao Zhang Haiteng Deng Jing Shi Man Pan Yi‐Ming Li Lei Liu 《Angewandte Chemie (International ed. in English)》2020,59(32):13496-13501
Triazole‐based deubiquitylase (DUB)‐resistant ubiquitin (Ub) probes have recently emerged as effective tools for the discovery of Ub chain‐specific interactors in proteomic studies, but their structural diversity is limited. A new family of DUB‐resistant Ub probes is reported based on isopeptide‐N‐ethylated dimeric or polymeric Ub chains, which can be efficiently prepared by a one‐pot, ubiquitin‐activating enzyme (E1)‐catalyzed condensation reaction of recombinant Ub precursors to give various homotypic and even branched Ub probes at multi‐milligram scale. Proteomic studies using label‐free quantitative (LFQ) MS indicated that the isopeptide‐N‐ethylated Ub probes may complement the triazole‐based probes in the study of Ub interactome. Our study highlights the utility of modern protein synthetic chemistry to develop structurally and new families of tool molecules needed for proteomic studies. 相似文献
20.
Sylvain Aubry Stphane Pellet‐Rostaing Marc Lemaire Ren Faure 《Journal of heterocyclic chemistry》2006,43(1):139-148