Exploring the mechanism of selective noncovalent adduct protein probing mass spectrometry utilizing site-directed mutagenesis to examine ubiquitin

Analytical Chemistry
Zhenjiu LiuRyan R Julian

Abstract

Mass spectrometry (MS) is emerging as an additional tool for examining protein structure by way of experiments where structurally related mass changes induced in solution are subsequently detected in the gas phase. Selective noncovalent adduct protein probing (SNAPP) is a recent addition to this type of experiment. SNAPP utilizes noncovalent recognition of lysine residues with 18-crown-6 (18C6) to monitor changes in protein structure. It has been observed that the number of 18C6 adducts that attach to a protein is a function of the structure of the protein. The present work seeks to examine the underlying chemistry which controls the differential attachment of 18C6 to lysine by using ubiquitin as a model system. Ubiquitin is a small protein with a structure that has been well characterized by multiple techniques. Site-directed mutagenesis was used to create a series of ubiquitin mutants where the lysine residues were exchanged for asparagine one at a time. These mutants were then evaluated by SNAPP-MS to determine the relative contribution of each lysine as a binding site for 18C6. It was found that attachment of 18C6 is largely controlled by the strength of intramolecular interactions involving lysine residues. Salt bridges pr...Continue Reading

References

Apr 5, 1987·Journal of Molecular Biology·S Vijay-KumarW J Cook
Feb 1, 1994·Protein Science : a Publication of the Protein Society·Z S Hendsch, B Tidor
Mar 1, 1994·Proteins·P L WintrodeP L Privalov
Oct 6, 1998·Annual Review of Biochemistry·A Hershko, A Ciechanover
Nov 5, 1999·Journal of Molecular Biology·S Kumar, R Nussinov
Nov 21, 2000·Trends in Biochemical Sciences·C M Pickart
Jul 18, 2001·Journal of the American Chemical Society·C R BabuA J Wand
Sep 5, 2002·Mass Spectrometry Reviews·Igor A Kaltashov, Stephen J Eyles
Jun 26, 2003·Protein Science : a Publication of the Protein Society·Laurence LinsRobert Brasseur
Jul 23, 2003·Nature Biotechnology·Junmin PengSteven P Gygi
Aug 12, 2004·Chemical Reviews·H Jane Dyson, Peter E Wright
Nov 24, 2004·Current Opinion in Chemical Biology·Cecile M Pickart, David Fushman
Mar 3, 2005·Journal of Molecular Biology·Ryo KitaharaKazuyuki Akasaka
Oct 7, 2005·Mass Spectrometry Reviews·Thomas E Wales, John R Engen
Jun 2, 2006·Analytical Chemistry·Alexis NazabalRenato Zenobi
Jun 13, 2006·Journal of the American Society for Mass Spectrometry·Tony Ly, Ryan R Julian

❮ Previous
Next ❯

Citations

Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Qingyu SunRyan R Julian
Feb 3, 2012·Journal of the American Society for Mass Spectrometry·Yuping Zhou, Richard W Vachet
Feb 22, 2012·Journal of the American Society for Mass Spectrometry·Yuping Zhou, Richard W Vachet
Aug 29, 2012·Journal of the American Society for Mass Spectrometry·Yu Chen, M T Rodgers
Dec 19, 2012·Journal of the American Society for Mass Spectrometry·Jong Wha LeeHugh I Kim
Apr 1, 2011·The Journal of Physical Chemistry. B·Brian C BohrerDavid E Clemmer
Mar 26, 2013·International Journal of Mass Spectrometry·Benjamin N MooreRyan R Julian
Mar 10, 2016·Chemical Reviews·M T Rodgers, P B Armentrout
Aug 11, 2016·Journal of the American Society for Mass Spectrometry·James G BonnerRyan R Julian
Nov 3, 2015·Rapid Communications in Mass Spectrometry : RCM·Yulin Qi, Dietrich A Volmer
Feb 27, 2013·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Martin F Czar, Rebecca A Jockusch
Nov 6, 2014·Chemical Communications : Chem Comm·Yanli TianChongzhao Ran
Oct 15, 2019·International Journal of Mass Spectrometry·Lance E TalbertRyan R Julian
May 21, 2010·Analytical Chemistry·Kevin PagelCarol V Robinson
Mar 10, 2012·Journal of the American Chemical Society·Yu Chen, M T Rodgers
Aug 16, 2012·The Journal of Physical Chemistry. a·Corey N StedwellNicolas C Polfer
Jan 14, 2012·Journal of the American Chemical Society·Yu Chen, M T Rodgers

❮ 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.