Protein and proteome phosphorylation stoichiometry analysis by element mass spectrometry

Analytical Chemistry
Ralf KrügerWolf D Lehmann

Abstract

Protein phosphorylation stoichiometry was assessed by two analytical strategies. Both are based on element mass spectrometry (ICPMS, inductively coupled plasma mass spectrometry) and simultaneous monitoring of (31)P and (34)S. One strategy employs a combination of 1D gel electrophoresis, in-gel digestion, and final microLC-ICPMS analysis (microLC = capillary liquid chromatography). The other strategy uses the combination of 1D gel electrophoresis, protein blotting, and imLA-ICPMS (imLA = imaging laser ablation). The two methods were evaluated with standard phosphoproteins and were applied to the analysis of the cytoplasmatic proteome of bacterial cells (Corynebacterium glutamicum) and eukaryotic cells (Mus musculus). The eukaryotic proteome was found to exhibit a significantly higher phosphorylation degree (approximately 0.8 mol of P/mol of protein) compared to the bacterial proteome (approximately 0.01 mol of P/mol of protein). Both analytical strategies revealed consistent quantitative results, with the microLC-ICPMS approach providing the higher sensitivity. In summary, two ICPMS-based methods for quantitative estimation of the phosphorylation degree of a cellular proteome are presented which access the native proteome state...Continue Reading

References

Jun 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·W J HenzelC Watanabe
May 18, 2001·Nature·P Blume-Jensen, T Hunter
May 4, 2002·Nature Cell Biology·Philip Cohen
Dec 10, 2002·Science·G ManningS Sudarsanam
Jan 24, 2003·Proceedings of the National Academy of Sciences of the United States of America·Debora BonenfantPaul Jenoe
Aug 19, 2003·Proteomics·Anne K BendtThomas Hermann
Sep 5, 2003·Journal of the American Society for Mass Spectrometry·William J HenzelJohn T Stults
Mar 23, 2004·Mini Reviews in Medicinal Chemistry·Eric C PetersScott B Ficarro
Mar 3, 2005·Proceedings of the National Academy of Sciences of the United States of America·Hanno SteenMarc W Kirschner
Mar 25, 2005·Nature Methods·Sam A Johnson, Tony Hunter
Apr 2, 2005·Analytical Chemistry·Jeffrey C SilvaScott Geromanos

❮ Previous
Next ❯

Citations

Aug 23, 2006·Analytical and Bioanalytical Chemistry·J BettmerA Prange
Nov 7, 2007·Analytical and Bioanalytical Chemistry·Christina Rappel, Dirk Schaumlöffel
Oct 17, 2008·Analytical and Bioanalytical Chemistry·Maria S JiménezJuan R Castillo
Mar 28, 2012·Journal of the American Society for Mass Spectrometry·Brittany CatronPatrick A Limbach
Dec 1, 2007·Chemical Communications : Chem Comm·Kagan Kerman, Heinz-Bernhard Kraatz
Mar 24, 2007·Clinical Chemistry and Laboratory Medicine : CCLM·Martin Kussmann
Oct 3, 2007·Analytical and Bioanalytical Chemistry·Alfredo Sanz-MedelJörg Bettmer
Jan 21, 2014·Current Opinion in Biotechnology·Aaron S Gajadhar, Forest M White
Aug 21, 2014·Mass Spectrometry Reviews·Evelyne MaesInge Mertens
May 28, 2009·Journal of Proteomics·Jörg BettmerAlfredo Sanz Medel
Dec 6, 2007·Journal of Trace Elements in Medicine and Biology : Organ of the Society for Minerals and Trace Elements (GMS)·Dirk Schaumlöffel
Feb 10, 2007·Biochemical and Biophysical Research Communications·Ralf KrügerWolf D Lehmann
Oct 12, 2010·Mass Spectrometry Reviews·Matthias TrostPierre Thibault
Jun 6, 2009·Mass Spectrometry Reviews·Meng WangZhi-Fang Chai
Nov 19, 2011·FEMS Microbiology Reviews·Ivan Mijakovic, Boris Macek
Apr 22, 2015·Biosensors & Bioelectronics·Nikhil BhallaPedro Estrela
Nov 29, 2011·Current Opinion in Biotechnology·Abderahmane DerouicheIvan Mijakovic
Jan 23, 2016·FEMS Microbiology Reviews·Amber J ParkCezar M Khursigara
Mar 23, 2017·GigaScience·Panayotis VlastaridisGrigoris D Amoutzias
Dec 1, 2006·Journal of Mass Spectrometry : JMS
Mar 28, 2007·Journal of Mass Spectrometry : JMS·Adrian A Ammann
Aug 3, 2007·Journal of Environmental Monitoring : JEM·Michael Krachler
Apr 28, 2020·FEBS Letters·Julie Bonne KøhlerIvan Mijakovic
Aug 26, 2020·Journal of the American Society for Mass Spectrometry·Julia A BubisFrank Kjeldsen

❮ 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.