scholarly journals Activated molecular probes for enzyme recognition and detection

Theranostics ◽  
2022 ◽  
Vol 12 (3) ◽  
pp. 1459-1485
Author(s):  
Meng Yuan ◽  
Ying Wu ◽  
Caiyan Zhao ◽  
Zhongxiang Chen ◽  
Lichao Su ◽  
...  
2014 ◽  
Vol 289 (44) ◽  
pp. 30556-30566 ◽  
Author(s):  
Lina Song ◽  
Collin Bachert ◽  
Katrine T. Schjoldager ◽  
Henrik Clausen ◽  
Adam D. Linstedt

Humans express up to 20 isoforms of GalNAc-transferase (herein T1–T20) that localize to the Golgi apparatus and initiate O-glycosylation. Regulation of this enzyme family affects a vast array of proteins transiting the secretory pathway and diseases arise upon misregulation of specific isoforms. Surprisingly, molecular probes to monitor GalNAc-transferase activity are lacking and there exist no effective global or isoform-specific inhibitors. Here we describe the development of T2- and T3-isoform specific fluorescence sensors that traffic in the secretory pathway. Each sensor yielded little signal when glycosylated but was strongly activated in the absence of its glycosylation. Specificity of each sensor was assessed in HEK cells with either the T2 or T3 enzymes deleted. Although the sensors are based on specific substrates of the T2 and T3 enzymes, elements in or near the enzyme recognition sequence influenced their activity and required modification, which we carried out based on previous in vitro work. Significantly, the modified T2 and T3 sensors were activated only in cells lacking their corresponding isozymes. Thus, we have developed T2- and T3-specific sensors that will be valuable in both the study of GalNAc-transferase regulation and in high-throughput screening for potential therapeutic regulators of specific GalNAc-transferases.


2014 ◽  
Vol 21 (21) ◽  
pp. 2367-2391 ◽  
Author(s):  
A. Gautam ◽  
P. Kapoor ◽  
K. Chaudhary ◽  
R. Kumar ◽  
Open Drug Discovery Consortium ◽  
...  

2013 ◽  
Vol 17 (6) ◽  
pp. 564-579 ◽  
Author(s):  
Junjia Liu ◽  
Chen Liu ◽  
Wei He

2014 ◽  
Vol 14 (1) ◽  
pp. 123-128 ◽  
Author(s):  
R. L. Mancinelli

AbstractWe have shown using ESA's Biopan facility flown in Earth orbit that when exposed to the space environment for 2 weeks the survival rate ofSynechococcus(Nägeli), a halophilic cyanobacterium isolated from the evaporitic gypsum–halite crusts that form along the marine intertidal, andHalorubrum chaoviatora member of the Halobacteriaceae isolated from an evaporitic NaCl crystal obtained from a salt evaporation pond, were higher than all other test organisms exceptBacillusspores. These results led to the EXPOSE-R mission to extend and refine these experiments as part of the experimental package for the external platform space exposure facility on the ISS. The experiment was flown in February 2009 and the organisms were exposed to low-Earth orbit for nearly 2 years. Samples were either exposed to solar ultraviolet (UV)-radiation (λ > 110 nm or λ > 200 nm, cosmic radiation (dosage range 225–320 mGy), or kept in darkness shielded from solar UV-radiation. Half of each of the UV-radiation exposed samples and dark samples were exposed to space vacuum and half kept at 105pascals in argon. Duplicate samples were kept in the laboratory to serve as unexposed controls. Ground simulation control experiments were also performed. After retrieval, organism viability was tested using Molecular Probes Live–Dead Bac-Lite stain and by their reproduction capability. Samples kept in the dark, but exposed to space vacuum had a 90 ± 5% survival rate compared to the ground controls. Samples exposed to full UV-radiation for over a year were bleached and although results from Molecular Probes Live–Dead stain suggested ~10% survival, the data indicate that no survival was detected using cell growth and division using the most probable number method. Those samples exposed to attenuated UV-radiation exhibited limited survival. Results from of this study are relevant to understanding adaptation and evolution of life, the future of life beyond earth, the potential for interplanetary transfer of viable microbes via meteorites and dust particles as well as spacecraft, and the physiology of halophiles.


1994 ◽  
Vol 269 (44) ◽  
pp. 27657-27662 ◽  
Author(s):  
D A Lawrence ◽  
S T Olson ◽  
S Palaniappan ◽  
D Ginsburg

2011 ◽  
Vol 54 (7) ◽  
pp. 2331-2340 ◽  
Author(s):  
Changning Wang ◽  
Chunying Wu ◽  
Junqing Zhu ◽  
Robert H. Miller ◽  
Yanming Wang

Molecules ◽  
2021 ◽  
Vol 26 (2) ◽  
pp. 485
Author(s):  
Veronika Huntosova ◽  
Denis Horvath ◽  
Robert Seliga ◽  
Georges Wagnieres

Detection of tissue and cell oxygenation is of high importance in fundamental biological and in many medical applications, particularly for monitoring dysfunction in the early stages of cancer. Measurements of the luminescence lifetimes of molecular probes offer a very promising and non-invasive approach to estimate tissue and cell oxygenation in vivo and in vitro. We optimized the evaluation of oxygen detection in vivo by [Ru(Phen)3]2+ in the chicken embryo chorioallantoic membrane model. Its luminescence lifetimes measured in the CAM were analyzed through hierarchical clustering. The detection of the tissue oxygenation at the oxidative stress conditions is still challenging. We applied simultaneous time-resolved recording of the mitochondrial probe MitoTrackerTM OrangeCMTMRos fluorescence and [Ru(Phen)3]2+ phosphorescence imaging in the intact cell without affecting the sensitivities of these molecular probes. [Ru(Phen)3]2+ was demonstrated to be suitable for in vitro detection of oxygen under various stress factors that mimic oxidative stress: other molecular sensors, H2O2, and curcumin-mediated photodynamic therapy in glioma cancer cells. Low phototoxicities of the molecular probes were finally observed. Our study offers a high potential for the application and generalization of tissue oxygenation as an innovative approach based on the similarities between interdependent biological influences. It is particularly suitable for therapeutic approaches targeting metabolic alterations as well as oxygen, glucose, or lipid deprivation.


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