Systematic characterization of phosphorylation sites in NFATc2 by linear ion trap mass spectrometry

Proteomics
Margarita VillarJesús Vázquez

Abstract

Members of the nuclear factor of activated T cells (NFAT) family of transcription factors regulate transcription of genes involved in the function of many different cellular systems. Activity of NFAT proteins is regulated by a complex interplay of phosphorylation events that are still poorly understood. In this work, we take advantage of the high scanning speed of the linear ion trap to develop a method to make a systematic characterization of NFATc2 phosphorylation. The method is based on the simultaneous monitoring of all tryptic peptides that can be detected by MS and contain potential phosphorylation sites. By this approach, we detected six NFATc2 phosphorylation sites by c-Jun NH2-terminal kinase (JNK) in vitro in only one experiment; a further site was also identified by performing digestion in solution. Using this approach, we have also characterized five basal phosphorylation sites in NFATc2 protein expressed in HEK cell cultures. Two of these NFATc2 phosphorylation sites in vivo have not been described before. The simplicity and sensitivity of this technique, which can be applied to any potential modification, makes it particularly attractive for the systematic detection of post-translational modifications of specific ...Continue Reading

References

Nov 1, 1984·Biomedical Mass Spectrometry·P Roepstorff, J Fohlman
Feb 1, 1996·Analytical Chemistry·M WilmM Mann
Dec 10, 1996·Proceedings of the National Academy of Sciences of the United States of America·A ShevchenkoM Mann
Jul 4, 1998·Journal of Chromatography. a·J X YanK L Williams
May 2, 2000·The Journal of Biological Chemistry·P Gómez del ArcoJ M Redondo
Apr 3, 2001·Nature Biotechnology·H ZhouR Aebersold
Feb 2, 2002·The Journal of Immunology : Official Journal of the American Association of Immunologists·Eeva-Marja RainioPäivi J Koskinen
Mar 5, 2002·Nature Biotechnology·Scott B FicarroForest M White
May 2, 2002·Cell·Gerald R Crabtree, Eric N Olson
May 15, 2002·Trends in Biotechnology·Matthias MannAkhilesh Pandey
Dec 10, 2002·Science·G ManningS Sudarsanam
Apr 15, 2003·Nature Biotechnology·Christine C WuJohn R Yates
Sep 17, 2003·Genes & Development·Patrick G HoganAnjana Rao
Aug 11, 2004·Proceedings of the National Academy of Sciences of the United States of America·Sean A BeausoleilSteven P Gygi
Sep 4, 2004·Molecular & Cellular Proteomics : MCP·Bryan A BallifSteven P Gygi
Mar 4, 2005·The Journal of Biological Chemistry·Inmaculada Ortega-PérezJuan Miguel Redondo
Jun 7, 2005·Proceedings of the National Academy of Sciences of the United States of America·Antonio Martínez-RuizSantiago Lamas

❮ Previous
Next ❯

Citations

Sep 3, 2010·Molecular & Cellular Proteomics : MCP·Elena Bonzon-KulichenkoJesús Vázquez
Dec 25, 2012·Applied and Environmental Microbiology·Leticia L TorresAurelio Hidalgo
Jan 29, 2010·Cardiovascular Research·Mariano Redondo-HorcajoSantiago Lamas
Oct 28, 2010·Biochimica Et Biophysica Acta·Cristina Díaz-JullienManuel Freire
Sep 15, 2009·Experimental Neurology·Massimo TabatonLuca Giliberto
Feb 24, 2009·Protein Expression and Purification·Juan Casado-VelaJ Ignacio Casal
Feb 21, 2009·Journal of Molecular Recognition : JMR·Tao LiuShaojun Liu
Aug 26, 2006·Trends in Immunology·Edgar SerflingAlois Palmetshofer
Jun 27, 2013·The Journal of Antimicrobial Chemotherapy·Federica CrucianiBeatrice Vitali

❮ 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.