A novel system to rapidly detect protein-protein interactions (PPIs) based on fluorescence co-localization.

Biotechnology Letters
Nan HuMin-Hui Pan

Abstract

Rapid and convenient detection of protein-protein interactions (PPIs) is of great significance for understanding function of protein. For efficiently detecting PPIs, we used the changes of proteins fluorescence localization to design a novel system, fluorescence translocation co-localization (FTCL), based on nuclear localization signal (NLS) in living cells. Depending on the original state of protein localization (both in the cytoplasm, both in the nucleus, one in the nucleus and another in the cytoplasm), two target proteins can be partitioned into the cytoplasm and nucleus by adding a NLS or mutating an existing NLS. Three independent results display that the changes of protein fluorescence co-localization were observed following co-expression of the two target proteins. At the same time, we verified the accuracy of fluorescence co-localization by co-immunoprecipitation. There FTCL system provided a novel detection method for PPIs, regardless of protein localization in the nucleus or cytoplasm. More importantly, this study provides a new strategy for future protein interaction studies through organelle localization (such as mitochondria, Golgi and cytomembrane, etc.).

References

Jun 14, 2001·Methods : a Companion to Methods in Enzymology·A J Walhout, M Vidal
Jun 20, 2001·Nature Reviews. Molecular Cell Biology·A J Walhout, M Vidal
Mar 5, 2003·The Journal of Cell Biology·Rajesh Babu Sekar, Ammasi Periasamy
Nov 8, 2003·Science·L GiotJ M Rothberg
Jan 6, 2004·Science·Siming LiMarc Vidal
Aug 12, 2005·Proceedings of the National Academy of Sciences of the United States of America·John P MillerStanley Fields
May 3, 2006·The American Journal of Pathology·Charles WalkerBen Low
Jul 4, 2006·Biophysical Journal·Huanmian ChenStephen R Ikeda
Apr 21, 2012·Journal of Proteome Research·Martine BeaufourHélène Bénédetti
Jun 20, 2012·Proteomics·Pascal Braun, Anne-Claude Gingras
Aug 22, 2012·Briefings in Functional Genomics·Javier De Las Rivas, Celia Fontanillo
Apr 2, 2017·Current Opinion in Structural Biology·Yoichi MurakamiKenji Mizuguchi
Oct 17, 2017·Methods in Molecular Biology·Henna IqbalMelisha R Kenedy
Feb 1, 2018·The Journal of Cell Biology·Sandra BackesJohannes M Herrmann
Apr 4, 2018·Accounts of Chemical Research·Rebecca FreilichJason E Gestwicki
Dec 5, 2019·Current Protocols in Toxicology·Yuekun LangHongmin Li

❮ Previous
Next ❯

Datasets Mentioned

BETA
AK406057.1

Methods Mentioned

BETA
ubiquitination
two-hybrid
pull-down
co-immunoprecipitation
immunoprecipitation
bimolecular fluorescence complementation
fluorescence resonance
fluorescence translocation
PCR
Fluorescence

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.