共查询到20条相似文献,搜索用时 31 毫秒
1.
Sam Benson Antonio Fernandez Nicole D. Barth Fabio deMoliner Mathew H. Horrocks C. Simon Herrington Jose Luis Abad Antonio Delgado Lisa Kelly Ziyuan Chang Yi Feng Miyako Nishiura Yuichiro Hori Kazuya Kikuchi Marc Vendrell 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2019,131(21):6985-6989
The transport and trafficking of metabolites are critical for the correct functioning of live cells. However, in situ metabolic imaging studies are hampered by the lack of fluorescent chemical structures that allow direct monitoring of small metabolites under physiological conditions with high spatial and temporal resolution. Herein, we describe SCOTfluors as novel small‐sized multi‐colored fluorophores for real‐time tracking of essential metabolites in live cells and in vivo and for the acquisition of metabolic profiles from human cancer cells of variable origin. 相似文献
2.
A Vinyl Sulfone‐Based Fluorogenic Probe Capable of Selective Labeling of PHGDH in Live Mammalian Cells 下载免费PDF全文
Dr. Sijun Pan Se‐Young Jang Si Si Liew Jiaqi Fu Danyang Wang Prof. Dr. Jun‐Seok Lee Prof. Dr. Shao Q. Yao 《Angewandte Chemie (International ed. in English)》2018,57(2):579-583
Chemical probes are powerful tools for interrogating small molecule‐target interactions. With additional fluorescence Turn‐ON functionality, such probes might enable direct measurements of target engagement in live mammalian cells. DNS‐pE (and its terminal alkyne‐containing version DNS‐pE2) is the first small molecule that can selectively label endogenous 3‐phosphoglycerate dehydrogenase (PHGDH) from various mammalian cells. Endowed with an electrophilic vinyl sulfone moiety that possesses fluorescence‐quenching properties, DNS‐pE/DNS‐pE2 became highly fluorescent only upon irreversible covalent modification of PHGDH. With an inhibitory property (in vitro Ki=7.4 μm ) comparable to that of known PHGDH inhibitors, our probes thus offer a promising approach to simultaneously image endogenous PHGDH activities and study its target engagement in live‐cell settings. 相似文献
3.
A Small‐Molecule FRET Reporter for the Real‐Time Visualization of Cell‐Surface Proteolytic Enzyme Functions 下载免费PDF全文
Jing Mu Dr. Fang Liu Muhammad Shafiq Rajab Meng Shi Shuang Li Chiching Goh Prof. Lei Lu Prof. Qing‐Hua Xu Prof. Bin Liu Dr. Lai Guan Ng Prof. Bengang Xing 《Angewandte Chemie (International ed. in English)》2014,53(52):14357-14362
Real‐time imaging of cell‐surface‐associated proteolytic enzymes is critical to better understand their performances in both physiological and pathological processes. However, most current approaches are limited by their complexity and poor membrane‐anchoring properties. Herein, we have designed and synthesized a unique small‐molecule fluorescent probe, which combines the principles of passive exogenous membrane insertion and Förster resonance energy transfer (FRET) to image cell‐surface‐localized furin‐like convertase activities. The membrane‐associated furin‐like enzymatic cleavage of the peptide probe leads to an increased fluorescence intensity which was mainly localized on the plasma membrane of the furin‐expressed cells. This small‐molecule fluorescent probe may serve as a unique and reliable reporter for real‐time visualization of endogenous cell‐surfaceassociated proteolytic furin‐like enzyme functions in live cells and tissues using one‐photon and two‐photon microscopy. 相似文献
4.
Xin Mao Dr. Peiyan Yuan Prof. Dr. Changmin Yu Prof. Dr. Lin Li Prof. Dr. Shao Q. Yao 《Angewandte Chemie (International ed. in English)》2018,57(32):10257-10262
Changes in the cellular levels of glutathione (GSH) and protein S‐glutathionylation (PSSG) are closely associated with a number of human diseases. Despite recent advances, few thiol‐reactive, small‐molecule GSH sensors could selectively detect GSH over other endogenous thiols, and none was capable of detecting PSSG in live mammalian cells. By using a dye‐loaded mesoporous silica nanoquencher (qMSN) capped with anti‐GSH antibody capable of highly selective binding toward GSH and glutathionylated proteins over other molecules, we have successfully developed a fluorescence GSH/PSSG nanosensor, which showed unprecedented selectivity toward PSSG even in the presence of GSH, had amplifiable and programmable fluorescence Turn‐ON properties, and could be used to image endogenous PSSG in live mammalian cells under stimulated conditions for the first time. 相似文献
5.
Molecular Imaging of Biological Samples on Nanophotonic Laser Desorption Ionization Platforms 下载免费PDF全文
Sylwia A. Stopka Charles Rong Dr. Andrew R. Korte Dr. Sridevi Yadavilli Dr. Javad Nazarian Dr. Trust T. Razunguzwa Dr. Nicholas J. Morris Prof. Akos Vertes 《Angewandte Chemie (International ed. in English)》2016,55(14):4482-4486
Mass spectrometry imaging (MSI) is a comprehensive tool for the analysis of a wide range of biomolecules. The mainstream method for molecular MSI is matrix‐assisted laser desorption ionization, however, the presence of a matrix results in spectral interferences and the suppression of some analyte ions. Herein we demonstrate a new matrix‐free MSI technique using nanophotonic ionization based on laser desorption ionization (LDI) from a highly uniform silicon nanopost array (NAPA). In mouse brain and kidney tissue sections, the distributions of over 80 putatively annotated molecular species are determined with 40 μm spatial resolution. Furthermore, NAPA‐LDI‐MS is used to selectively analyze metabolites and lipids from sparsely distributed algal cells and the lamellipodia of human hepatocytes. Our results open the door for matrix‐free MSI of tissue sections and small cell populations by nanophotonic ionization. 相似文献
6.
Dinuclear Ruthenium(II) Complexes as Two‐Photon,Time‐Resolved Emission Microscopy Probes for Cellular DNA 下载免费PDF全文
Elizabeth Baggaley Martin R. Gill Nicola H. Green David Turton Igor V. Sazanovich Stanley W. Botchway Carl Smythe John W. Haycock Julia A. Weinstein Jim A. Thomas 《Angewandte Chemie (International ed. in English)》2014,53(13):3367-3371
The first transition‐metal complex‐based two‐photon absorbing luminescence lifetime probes for cellular DNA are presented. This allows cell imaging of DNA free from endogenous fluorophores and potentially facilitates deep tissue imaging. In this initial study, ruthenium(II) luminophores are used as phosphorescent lifetime imaging microscopy (PLIM) probes for nuclear DNA in both live and fixed cells. The DNA‐bound probes display characteristic emission lifetimes of more than 160 ns, while shorter‐lived cytoplasmic emission is also observed. These timescales are orders of magnitude longer than conventional FLIM, leading to previously unattainable levels of sensitivity, and autofluorescence‐free imaging. 相似文献
7.
Raman Microspectroscopic Evidence for the Metabolism of a Tyrosine Kinase Inhibitor,Neratinib, in Cancer Cells 下载免费PDF全文
Karim Aljakouch Tatjana Lechtonen Dr. Hesham K. Yosef Mohamad K. Hammoud Wissam Alsaidi Dr. Carsten Kötting Dr. Carolin Mügge Prof. Dr. Robert Kourist Dr. Samir F. El‐Mashtoly Prof. Dr. Klaus Gerwert 《Angewandte Chemie (International ed. in English)》2018,57(24):7250-7254
Tyrosine kinase receptors are one of the main targets in cancer therapy. They play an essential role in the modulation of growth factor signaling and thereby inducing cell proliferation and growth. Tyrosine kinase inhibitors such as neratinib bind to EGFR and HER2 receptors and exhibit antitumor activity. However, little is known about their detailed cellular uptake and metabolism. Here, we report for the first time the intracellular spatial distribution and metabolism of neratinib in different cancer cells using label‐free Raman imaging. Two new neratinib metabolites were detected and fluorescence imaging of the same cells indicate that neratinib accumulates in lysosomes. The results also suggest that both EGFR and HER2 follow the classical endosome lysosomal pathway for degradation. A combination of Raman microscopy, DFT calculations, and LC‐MS was used to identify the chemical structure of neratinib metabolites. These results show the potential of Raman microscopy to study drug pharmacokinetics. 相似文献
8.
Jae Won Chang Mohammed Bhuiyan Hsiu‐Ming Tsai Hannah J. Zhang Gang Li Shaghayegh Fathi David C. McCutcheon Lara Leoni Richard Freifelder Chin‐Tu Chen Raymond E. Moellering 《Angewandte Chemie (International ed. in English)》2020,59(35):15161-15165
Herein, we report the development of an 18F‐labeled, activity‐based small‐molecule probe targeting the cancer‐associated serine hydrolase NCEH1. We undertook a focused medicinal chemistry campaign to simultaneously preserve potent and specific NCEH1 labeling in live cells and animals, while permitting facile 18F radionuclide incorporation required for PET imaging. The resulting molecule, [18F]JW199, labels active NCEH1 in live cells at nanomolar concentrations and greater than 1000‐fold selectivity relative to other serine hydrolases. [18F]JW199 displays rapid, NCEH1‐dependent accumulation in mouse tissues. Finally, we demonstrate that [18F]JW199 labels aggressive cancer tumor cells in vivo, which uncovered localized NCEH1 activity at the leading edge of triple‐negative breast cancer tumors, suggesting roles for NCEH1 in tumor aggressiveness and metastasis. 相似文献
9.
Philipp Werther Klaus Yserentant Felix Braun Nicolai Kaltwasser Christoph Popp Mathis Baalmann Dirk‐Peter Herten Richard Wombacher 《Angewandte Chemie (International ed. in English)》2020,59(2):804-810
Recent developments in fluorescence microscopy call for novel small‐molecule‐based labels with multiple functionalities to satisfy different experimental requirements. A current limitation in the advancement of live‐cell single‐molecule localization microscopy is the high excitation power required to induce blinking. This is in marked contrast to the minimal phototoxicity required in live‐cell experiments. At the same time, quality of super‐resolution imaging depends on high label specificity, making removal of excess dye essential. Approaching both hurdles, we present the design and synthesis of a small‐molecule label comprising both fluorogenic and self‐blinking features. Bioorthogonal click chemistry ensures fast and highly selective attachment onto a variety of biomolecular targets. Along with spectroscopic characterization, we demonstrate that the probe improves quality and conditions for regular and single‐molecule localization microscopy on live‐cell samples. 相似文献
10.
Xiu‐Cai Chen Dr. Shuo‐Bin Chen Jing Dai Jia‐Hao Yuan Prof. Tian‐Miao Ou Prof. Zhi‐Shu Huang Prof. Jia‐Heng Tan 《Angewandte Chemie (International ed. in English)》2018,57(17):4702-4706
Because of the absence of methods for tracking RNA G‐quadruplex dynamics, especially the folding and unfolding of this attractive structure in live cells, understanding of the biological roles of RNA G‐quadruplexes is so far limited. Herein, we report a new red‐emitting fluorescent probe, QUMA‐1 , for the selective, continuous, and real‐time visualization of RNA G‐quadruplexes in live cells. The applications of QUMA‐1 in several previously intractable applications, including live‐cell imaging of the dynamic folding, unfolding, and movement of RNA G‐quadruplexes and the visualization of the unwinding of RNA G‐quadruplexes by RNA helicase have been demonstrated. Notably, our real‐time results revealed the complexity of the dynamics of RNA G‐quadruplexes in live cells. We anticipate that the further application of QUMA‐1 in combination with appropriate biological and imaging methods to explore the dynamics of RNA G‐quadruplexes will uncover more information about the biological roles of RNA G‐quadruplexes. 相似文献
11.
Dr. Shin‐ichi Sato Dr. Mizuki Watanabe Dr. Yousuke Katsuda Dr. Asako Murata Dr. Dan Ohtan Wang Prof. Motonari Uesugi 《Angewandte Chemie (International ed. in English)》2015,54(6):1855-1858
Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. A new convenient and versatile method is reported that permits spatiotemporal imaging of specific non‐engineered RNAs in living cells. The method uses transfection of a plasmid encoding a gene‐specific RNA aptamer, combined with a cell‐permeable synthetic small molecule, the fluorescence of which is restored only when the RNA aptamer hybridizes with its cognitive mRNA. The method was validated by live‐cell imaging of the endogenous mRNA of β‐actin. Application of the technology to mRNAs of a total of 84 human cytoskeletal genes allowed us to observe cellular dynamics of several endogenous mRNAs including arfaptin‐2, cortactin, and cytoplasmic FMR1‐interacting protein 2. The RNA‐imaging technology and its further optimization might permit live‐cell imaging of any RNA molecules. 相似文献
12.
Sandra Resa Dr. Angel Orte Dr. Delia Miguel Dr. Jose M. Paredes Virginia Puente‐Muñoz Dr. Rafael Salto Dr. Maria D. Giron Dr. Maria J. Ruedas‐Rama Dr. Juan M. Cuerva Prof. Jose M. Alvarez‐Pez Dr. Luis Crovetto 《Chemistry (Weinheim an der Bergstrasse, Germany)》2015,21(42):14772-14779
The simultaneous detection of relevant metabolites in living organisms by using one molecule introduces an approach to understanding the relationships between these metabolites in healthy and deregulated cells. Fluorescent probes of low toxicity are remarkable tools for this type of analysis of biological systems in vivo. As a proof of concept, different naturally occurring compounds, such as biothiols and phosphate anions, were the focus for this work. The 2,4‐dinitrobenzenesulfinate (DNBS) derivative of 9‐[1‐(4‐tert‐butyl‐2‐methoxyphenyl)]‐6‐hydroxy‐3H‐xanthen‐3‐one (Granada Green; GG) were designed and synthesized. This new sulfinyl xanthene derivative can act as a dual sensor for the aforementioned analytes simultaneously. The mechanism of action of this derivative implies thiolysis of the sulfinyl group of the weakly fluorescent DNBS‐GG by biological thiols at near‐neutral pH values, thus releasing the fluorescent GG moiety, which simultaneously responds to phosphate anions through its fluorescence‐decay time. The new dual probe was tested in solution by using steady‐state and time‐resolved fluorescence and intracellularly by using fluorescence‐lifetime imaging microscopy (FLIM) in human epithelioid cervix carcinoma (HeLa) cells. 相似文献
13.
Far‐red organic fluorophores commonly used in traditional and super‐resolution localization microscopy are found to contain a fluorescent impurity with green excitation and near‐red emission. This near‐red fluorescent impurity can interfere with some multicolor stochastic optical reconstruction microscopy/photoactivated localization microscopy measurements in live cells and produce subtle artifacts in chemically fixed cells. We additionally describe alternatives to avoid artifacts in super‐resolution localization microscopy. 相似文献
14.
In Situ Synthesis of Alkenyl Tetrazines for Highly Fluorogenic Bioorthogonal Live‐Cell Imaging Probes 下载免费PDF全文
Dr. Haoxing Wu Dr. Jun Yang Dr. Jolita Šečkutė Prof. Neal K. Devaraj 《Angewandte Chemie (International ed. in English)》2014,53(23):5805-5809
In spite of the wide application potential of 1,2,4,5‐tetrazines, particularly in live‐cell and in vivo imaging, a major limitation has been the lack of practical synthetic methods. Here we report the in situ synthesis of (E)‐3‐substituted 6‐alkenyl‐1,2,4,5‐tetrazine derivatives through an elimination–Heck cascade reaction. By using this strategy, we provide 24 examples of π‐conjugated tetrazine derivatives that can be conveniently prepared from tetrazine building blocks and related halides. These include tetrazine analogs of biological small molecules, highly conjugated buta‐1,3‐diene‐substituted tetrazines, and a diverse array of fluorescent probes suitable for live‐cell imaging. These highly conjugated probes show very strong fluorescence turn‐on (up to 400‐fold) when reacted with dienophiles such as cyclopropenes and trans‐cyclooctenes, and we demonstrate their application for live‐cell imaging. This work provides an efficient and practical synthetic methodology for tetrazine derivatives and will facilitate the application of conjugated tetrazines, particularly as fluorogenic probes for live‐cell imaging. 相似文献
15.
Mapping Live Cell Viscosity with an Aggregation‐Induced Emission Fluorogen by Means of Two‐Photon Fluorescence Lifetime Imaging 下载免费PDF全文
Dr. Sijie Chen Dr. Yuning Hong Dr. Yan Zeng Qiqi Sun Dr. Yang Liu Engui Zhao Gongxun Bai Prof. Jianan Qu Prof. Jianhua Hao Prof. Ben Zhong Tang 《Chemistry (Weinheim an der Bergstrasse, Germany)》2015,21(11):4315-4320
Intracellular viscosity is a crucial parameter that indicates the functioning of cells. In this work, we demonstrate the utility of TPE‐Cy, a cell‐permeable dye with aggregation‐induced emission (AIE) property, in mapping the viscosity inside live cells. Owing to the AIE characteristics, both the fluorescence intensity and lifetime of this dye are increased along with an increase in viscosity. Fluorescence lifetime imaging of live cells stained with TPE‐Cy reveals that the lifetime in lipid droplets is much shorter than that from the general cytoplasmic region. The loose packing of the lipids in a lipid droplet results in low viscosity and thus shorter lifetime of TPE‐Cy in this region. It demonstrates that the AIE dye could provide good resolution in intracellular viscosity sensing. This is also the first work in which AIE molecules are applied in fluorescence lifetime imaging and intracellular viscosity sensing. 相似文献
16.
Dr. Zbigniew Zawada Dr. Ameneh Tatar Dr. Pavle Mocilac Dr. Miloš Buděšínský Dr. Tomáš Kraus 《Angewandte Chemie (International ed. in English)》2018,57(31):9891-9895
Chemically modified nucleoside triphosphates (NTPs) are widely exploited as unnatural metabolites in chemical biology and medicinal chemistry. Because anionic NTPs do not permeate cell membranes, their corresponding neutral precursors are employed in cell‐based assays. These precursors become active metabolites after enzymatic conversion, which often proceeds insufficiently. Here we show that metabolically‐active NTPs can be directly transported into eukaryotic cells and bacteria by the action of designed synthetic nucleoside triphosphate transporters (SNTTs). The transporter is composed of a receptor, which forms a non‐covalent complex with a triphosphate anion, and a cell‐penetrating agent, which translocates the complex across the plasma membrane. NTP is then released from the complex in the intracellular milieu and accumulates in nuclei and nucleoli in high concentration. The transport of NTPs proceeds rapidly (seconds to minutes) and selectively even in the presence of other organic anions. We demonstrate that this operationally simple and efficient means of transport of fluorescently labelled NTPs into cells can be used for metabolic labeling of DNA in live cells. 相似文献
17.
Rigumula Wu Aruni P. K. K. Karunanayake Mudiyanselage Fatemeh Shafiei Bin Zhao Yousef Bagheri Qikun Yu Kathleen McAuliffe Kewei Ren Mingxu You 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2019,131(50):18439-18443
Precisely determining the intracellular concentrations of metabolites and signaling molecules is critical in studying cell biology. Fluorogenic RNA‐based sensors have emerged to detect various targets in living cells. However, it is still challenging to apply these genetically encoded sensors to quantify the cellular concentrations and distributions of targets. Herein, using a pair of orthogonal fluorogenic RNA aptamers, DNB and Broccoli, we engineered a modular sensor system to apply the DNB‐to‐Broccoli fluorescence ratio to quantify the cell‐to‐cell variations of target concentrations. These ratiometric sensors can be broadly applied for live‐cell imaging and quantification of metabolites, signaling molecules, and other synthetic compounds. 相似文献
18.
A derivatization and validation strategy for determining the spatial localization of endogenous amine metabolites in tissues using MALDI imaging mass spectrometry 下载免费PDF全文
M. L. Manier J. M. Spraggins M. L. Reyzer J. L. Norris R. M. Caprioli 《Journal of mass spectrometry : JMS》2014,49(8):ii-ii
Imaging mass spectrometry (IMS) studies increasingly focus on endogenous small molecular weight metabolites and consequently bring special analytical challenges. Since analytical tissue blanks do not exist for endogenous metabolites, careful consideration must be given to confirm molecular identity. Here, we present approaches for the improvement in detection of endogenous amine metabolites such as amino acids and neurotransmitters in tissues through chemical derivatization and matrix‐assisted laser desorption/ionization (MALDI) IMS. Chemical derivatization with 4‐hydroxy‐3‐methoxycinnamaldehyde (CA) was used to improve sensitivity and specificity. CA was applied to the tissue via MALDI sample targets precoated with a mixture of derivatization reagent and ferulic acid as a MALDI matrix. Spatial distributions of chemically derivatized endogenous metabolites in tissue were determined by high‐mass resolution and MSn IMS. We highlight an analytical strategy for metabolite validation whereby tissue extracts are analyzed by high‐performance liquid chromatography (HPLC)‐MS/MS to unambiguously identify metabolites and distinguish them from isobaric compounds. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
19.
Firefly Luciferase Mutants Allow Substrate‐Selective Bioluminescence Imaging in the Mouse Brain 下载免费PDF全文
Spencer T. Adams Jr. Dr. David M. Mofford Dr. G. S. Kiran Kumar Reddy Prof. Stephen C. Miller 《Angewandte Chemie (International ed. in English)》2016,55(16):4943-4946
Bioluminescence imaging is a powerful approach for visualizing specific events occurring inside live mice. Animals can be made to glow in response to the expression of a gene, the activity of an enzyme, or the growth of a tumor. But bioluminescence requires the interaction of a luciferase enzyme with a small‐molecule luciferin, and its scope has been limited by the mere handful of natural combinations. Herein, we show that mutants of firefly luciferase can discriminate between natural and synthetic substrates in the brains of live mice. When using adeno‐associated viral (AAV) vectors to express luciferases in the brain, we found that mutant luciferases that are inactive or weakly active with d ‐luciferin can light up brightly when treated with the aminoluciferins CycLuc1 and CycLuc2 or their respective FAAH‐sensitive luciferin amides. Further development of selective luciferases promises to expand the power of bioluminescence and allow multiple events to be imaged in the same live animal. 相似文献
20.
A Cell‐Targeted Non‐Cytotoxic Fluorescent Nanogel Thermometer Created with an Imidazolium‐Containing Cationic Radical Initiator 下载免费PDF全文
Dr. Seiichi Uchiyama Dr. Toshikazu Tsuji Kyoko Kawamoto Dr. Kentaro Okano Eiko Fukatsu Takahiro Noro Kumiko Ikado Sayuri Yamada Yuka Shibata Dr. Teruyuki Hayashi Dr. Noriko Inada Dr. Masaru Kato Dr. Hideki Koizumi Prof. Hidetoshi Tokuyama 《Angewandte Chemie (International ed. in English)》2018,57(19):5413-5417
A cationic fluorescent nanogel thermometer based on thermo‐responsive N‐isopropylacrylamide and environment‐sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02–0.84 °C in the range 20–40 °C), and remarkable non‐cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells. 相似文献