Method for distinguishing specific from nonspecific protein-ligand complexes in nanoelectrospray ionization mass spectrometry

Analytical Chemistry
Jiangxiao SunJohn S Klassen

Abstract

A new methodology for distinguishing between specific and nonspecific protein-ligand complexes in nanoelectrospray ionization mass spectrometry (nanoES-MS) is described. The method involves the addition of an appropriate reference protein (P(ref)), which does not bind specifically to any of the solution components, to the nanoES solution containing the protein(s) and ligand(s) of interest. The occurrence of nonspecific protein-ligand binding is monitored by the appearance of nonspecific (P(ref) + ligand) complexes in the nanoES mass spectrum. Furthermore, the fraction of P(ref) undergoing nonspecific ligand binding provides a quantitative measure of the contribution of nonspecific binding to the measured intensities of protein and specific protein-ligand complexes. As a result, errors introduced into protein-ligand association constants, K(assoc), as determined with nanoES-MS, by nonspecific ligand binding can be corrected. The principal assumptions on which this methodology is based, namely, that the fraction of proteins and protein complexes that engage in nonspecific ligand binding during the nanoES process is determined by the number of free ligand molecules in the offspring droplets leading to gaseous ions and is independe...Continue Reading

References

Jan 1, 1994·Methods in Enzymology·C P GlaudemansE M Nashed
Mar 4, 2000·Journal of the American Society for Mass Spectrometry·K HåkanssonP Håkansson
Nov 23, 2000·Proceedings of the National Academy of Sciences of the United States of America·M FändrichC V Robinson
Sep 13, 2003·Methods in Enzymology·Kiyohito Shimura, Ken-ichi Kasai
Sep 13, 2003·Methods in Enzymology·Pernilla SörmeHakon Leffler
Apr 15, 2004·Journal of the American Chemical Society·Rambod DaneshfarJohn S Klassen

❮ Previous
Next ❯

Citations

Jan 25, 2012·Journal of the American Society for Mass Spectrometry·Elena N KitovaJohn S Klassen
Aug 15, 2013·Journal of the American Society for Mass Spectrometry·Yixuan ZhangJohn S Klassen
Jun 21, 2008·Journal of the American Society for Mass Spectrometry·Sheng YinJoseph A Loo
Mar 27, 2009·Journal of the American Society for Mass Spectrometry·Nian SunJohn S Klassen
Jan 22, 2010·Journal of the American Society for Mass Spectrometry·Nian SunJohn S Klassen
Jun 29, 2010·Journal of the American Society for Mass Spectrometry·Eric MartineauAhmed M Aman
Aug 31, 2010·Journal of the American Society for Mass Spectrometry·Amr El-HawietJohn S Klassen
Nov 17, 2012·Inorganic Chemistry·Amit S PithadiaMi Hee Lim
May 26, 2011·Glycobiology·Amr El-HawietJohn S Klassen
Mar 27, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Gillian R HiltonJustin L P Benesch
Oct 15, 2013·Journal of the American Society for Mass Spectrometry·Hong LinJohn S Klassen
Oct 8, 2013·Journal of the American Society for Mass Spectrometry·Ling HanJohn S Klassen
Aug 20, 2014·Journal of the American Society for Mass Spectrometry·Elena N KitovaJohn S Klassen
Dec 17, 2015·Journal of the American Society for Mass Spectrometry·Yuyu YaoJohn S Klassen
Feb 4, 2016·Journal of the American Chemical Society·Eric LargyJean-Louis Mergny
Nov 27, 2015·Protein Science : a Publication of the Protein Society·Mikko LaitaojaJanne Jänis
Jun 25, 2009·Mass Spectrometry Reviews·Paul Kebarle, Udo H Verkerk
Jul 31, 2013·Journal of Mass Spectrometry : JMS·C SantambrogioH Molinari
Aug 13, 2009·Mass Spectrometry Reviews·Lars KonermannXin Tong
May 28, 2014·Rapid Communications in Mass Spectrometry : RCM·Hannah J MapleRebecca J Burnley
May 15, 2015·Protein Science : a Publication of the Protein Society·Shuai Niu, Brandon T Ruotolo
May 6, 2015·Glycobiology·Amr El-HawietJohn S Klassen
May 9, 2015·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Federico Riccardi SirtoriGianpaolo Fogliatto
Oct 16, 2014·Journal of the American Society for Mass Spectrometry·Yuyu YaoJohn S Klassen
Jul 29, 2016·Analytical Chemistry·Albert KonijnenbergAntonino Natalello
Aug 24, 2016·Journal of the American Society for Mass Spectrometry·Ling HanJohn S Klassen
Dec 3, 2016·Journal of the American Society for Mass Spectrometry·Aneika C Leney, Albert J R Heck
Feb 1, 2017·Chembiochem : a European Journal of Chemical Biology·Kalli C CatcottZhaohui Sunny Zhou
Sep 5, 2017·Angewandte Chemie·Sem TamaraAneika C Leney

❮ 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.