Quantitative determination of protein stability and ligand binding by pulse proteolysis

Current Protocols in Protein Science
Chiwook Park, Susan Marqusee

Abstract

Pulse proteolysis exploits the difference in proteolytic susceptibility between folded and unfolded proteins for facile but quantitative determination of protein stability. The method requires only common biochemistry and molecular biology lab equipment. Pulse proteolysis also can be used to determine the affinity of a ligand to its protein target by monitoring the change in protein stability upon ligand binding. The Basic Protocol describes the detailed procedure for determining protein stability using pulse proteolysis. For pulse proteolysis to be used for determining a protein's stability, the protein should not be digested significantly by pulse proteolysis when it is in the folded conformation. The Support Protocol describes a procedure for determining whether a protein satisfies this requirement. The principles of protein stability determination using denaturant and pulse proteolysis are also discussed.

References

Mar 9, 1976·Biochemistry·F W DahlquistW L Bigbee
Nov 1, 1989·Analytical Biochemistry·S C Gill, P H von Hippel
Aug 20, 1986·Journal of Molecular Biology·T ImotoT Ueda
Oct 1, 1995·Protein Science : a Publication of the Protein Society·J K MyersJ M Scholtz
Oct 20, 2004·Journal of Molecular Biology·Chiwook Park, Susan Marqusee
Mar 10, 2005·Chemical Reviews·Joel D A TyndallDavid P Fairlie

❮ Previous
Next ❯

Citations

May 4, 2012·Biochemistry·Robin RoychaudhuriDavid B Teplow
Apr 24, 2016·Nucleic Acids Research·Brianne E DocterJames C A Bardwell
Apr 24, 2009·Protein Science : a Publication of the Protein Society·Moon-Soo KimChiwook Park
Feb 13, 2013·Protein Science : a Publication of the Protein Society·Jay H ChoiMarc Ostermeier
Jul 30, 2010·Protein Science : a Publication of the Protein Society·Melinda S HanesTracy M Handel
May 10, 2020·Protein Science : a Publication of the Protein Society·Chen Chen, Chiwook Park
Aug 9, 2012·The Journal of Biological Chemistry·Julie A ZornJames A Wells
Aug 13, 2013·G3 : Genes - Genomes - Genetics·Dago Dimster-DenkJasper Rine
Nov 19, 2011·Protein Science : a Publication of the Protein Society·Jonathan P SchlebachChiwook Park
Jun 13, 2020·The Journal of Biological Chemistry·Madeleine K JensenSusan Marqusee
Jan 9, 2013·The Journal of Biological Chemistry·Brian R AndersonHenk Granzier
Mar 6, 2018·Current Opinion in Structural Biology·Kritika Gupta, Raghavan Varadarajan
Jan 5, 2011·Journal of Molecular Biology·Jonathan P SchlebachChiwook Park
Feb 23, 2011·Journal of Molecular Biology·Pei-Fen LiuChiwook Park

❮ 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.