In situ, real-time monitoring of the 3' to 5' exonucleases secreted by living cells

Analytical Chemistry
Xin SuMeiping Zhao

Abstract

Enzymes containing 3'-5' exonuclease activities play vital roles in maintaining genome stability. Though a wide variety of methods have been developed for detection of these enzymes, few of them can be directly applied for in situ and real-time monitoring of the secretion of these active substances by living cells. Taking advantages of the free 3'-end of stacked guanine-quenched photoinduced electron transfer fluorescent probes, here we demonstrate a novel assay capable of in situ and real-time monitoring of the 3'-5' exonucleases secreted by living cells. The detection limit of the new method achieved as low as 0.04 U/mL, allowing direct monitoring of the target enzymes in an extracellular environment without preconcentration steps. False positive signals caused by other nonspecific enzymes were easily ruled out by the use of a control probe with the 3'-end modified with exonuclease-resistant phosphorothioate guanines. Using Alexa Fluor 488 as the fluorophore, the probe is adaptable to a wide range of pH conditions. The approach was successfully applied for in situ, real-time monitoring of the 3'-5' exonucleases secreted by suspension cells of Arabidopsis thaliana. It also holds great potential for in situ and real-time detect...Continue Reading

References

Jan 1, 1994·Annual Review of Biochemistry·B Demple, L Harrison
Sep 24, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·M GilarA S Cohen
Aug 17, 1999·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·N Panchuk-VoloshinaR P Haugland
May 4, 2002·Nature Reviews. Molecular Cell Biology·Igor V Shevelev, Ulrich Hübscher
Oct 5, 2002·Nucleic Acids Research·Andrew TsourkasGang Bao
Feb 4, 2011·Chemical Society Reviews·Nan Dai, Eric T Kool

❮ Previous
Next ❯

Citations

May 4, 2017·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·Dik-Lung MaChung-Hang Leung
May 18, 2013·International Journal of Molecular Sciences·Sha ZhuGenxi Li
Nov 14, 2012·Applied Spectroscopy·Xin SuMeiping Zhao
Feb 2, 2013·Chembiochem : a European Journal of Chemical Biology·Jong-Wha JungEric T Kool
Jan 21, 2015·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Dik-Lung MaChung-Hang Leung
Oct 14, 2014·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Modi WangDik-Lung Ma
Oct 17, 2019·Enzyme and Microbial Technology·Konstantin M BurkinIvan Yu Sakharov
Mar 11, 2021·Analytical Methods : Advancing Methods and Applications·Xu ZhangChengzhi Huang
May 10, 2013·Analytical Chemistry·Yibin LiuMeiping Zhao
Nov 22, 2012·Analytical Chemistry·Qinfeng XuChun-yang Zhang

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.