Development of a targeted selected ion monitoring assay for the elucidation of protease induced structural changes in cardiac troponin T

Journal of Proteomics
Alexander S StrengWill K W H Wodzig

Abstract

Cardiac troponin T (cTnT) is a highly cardiospecific protein commonly used in the diagnosis of acute myocardial infarction (AMI), but is subject to proteolytic degradation upon its release in the circulation. In this study, a targeted mass spectrometry assay was developed to detect peptides which are differentially present within the different degradation products. cTnT was spiked in human serum and incubated at 37 °C to induce proteolytic degradation. Isolation and fractionation of cTnT and its fragments from serum were performed using immunoprecipitation and SDS-PAGE. Bands migrating to 37 kDa (intact cTnT), 29 kDa (primary fragment), and 19, 18, and 16kDa (secondary fragments) were excised, digested, and subsequently analysed using targeted selected ion monitoring on a UHPLC-coupled quadrupole-Orbitrap mass spectrometer. Sixteen precursor ions from a total of 11 peptides unique to cTnT were targeted. Precursor ions were detectable up until 1200 ng/L cTnT, which is a typical cTnT concentration after AMI. With tandem-MS and relative quantification, we proved the formation of cTnT fragments upon incubation in human serum and identified differentially present peptides in the fragment bands, indicative of N- and C-terminal proteo...Continue Reading

References

Apr 25, 2002·Proceedings of the National Academy of Sciences of the United States of America·Catherine CommunalRoger J Hajjar
Jan 10, 2003·Nucleic Acids Research·Cathy H WuWinona C Barker
Mar 14, 2003·Nature·Ruedi Aebersold, Matthias Mann
Jun 16, 2005·Nature Reviews. Molecular Cell Biology·Bernhard KusterRuedi Aebersold
Apr 15, 2006·Science·Bruno Domon, Ruedi Aebersold
May 22, 2007·Clinical Biochemistry·Etienne C H J MichielsenMarja P Van Dieijen-Visser
Mar 14, 2008·Rapid Communications in Mass Spectrometry : RCM·Chiara CavaliereAldo Laganà
Oct 16, 2008·Molecular Systems Biology·Vinzenz LangeRuedi Aebersold
May 19, 2009·Nature Methods·Ruedi Aebersold
Jul 15, 2009·Molecular & Cellular Proteomics : MCP·Hasmik KeshishianSteven A Carr
Jul 14, 2010·Nature Biotechnology·Bruno Domon, Ruedi Aebersold
Oct 23, 2010·Archives of Biochemistry and Biophysics·Bin Wei, J-P Jin
Mar 12, 2011·Journal of Mass Spectrometry : JMS·Sebastien GallienBruno Domon
Aug 23, 2011·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mark S LowenthalDavid M Bunk
Aug 7, 2012·Molecular & Cellular Proteomics : MCP·Amelia C PetersonJoshua J Coon
Aug 11, 2012·Bioanalysis·Stephen W HolmanClaire E Eyers
Sep 11, 2012·Molecular & Cellular Proteomics : MCP·Sebastien GallienBruno Domon
Sep 11, 2012·Clinical Chemistry and Laboratory Medicine : CCLM·Alma M A MingelsMarja P van Dieijen-Visser
Dec 4, 2012·Nucleic Acids Research·Juan Antonio VizcaínoHenning Hermjakob
Dec 22, 2012·Analytical Chemistry·Sergei V SavelievMarjeta Urh
Mar 29, 2013·Clinical Chemistry·Eline P M CardinaelsMarja P van Dieijen-Visser
Jul 23, 2013·Journal of Molecular and Cellular Cardiology·Alexander S StrengWill K W H Wodzig
Jul 31, 2013·International Journal of Cardiology·Moritz BienerEvangelos Giannitsis
Aug 21, 2013·Bioinformatics·Chuming ChenUNKNOWN UniProt Consortium
Feb 5, 2014·Biochemistry. Biokhimii︠a︡·I A Katrukha
Mar 13, 2014·Experimental and Molecular Pathology·Alexander S StrengMarja P van Dieijen-Visser
Sep 26, 2014·Clinical Chemistry and Laboratory Medicine : CCLM·Petr Jarolim
Dec 1, 2012·Global Heart·Kristian ThygesenHarvey D White
Apr 1, 2015·The FEBS Journal·Liana Tsiatsiani, Albert J R Heck

❮ Previous
Next ❯

Citations

Nov 7, 2016·Biochemical and Biophysical Research Communications·Alexander S StrengWill K W H Wodzig
Feb 15, 2017·Proteomics. Clinical Applications·Somchai ChutipongtanateJisnuson Svasti
Jun 13, 2018·Annual Review of Analytical Chemistry·J Astor AnkneyXian Chen
Jul 7, 2018·Biomarkers : Biochemical Indicators of Exposure, Response, and Susceptibility to Chemicals·Ola HammarstenAllan S Jaffe
Apr 22, 2017·Clinical Chemistry·Geza S Bodor
Apr 22, 2017·Clinical Chemistry·Ivan A KatrukhaAlexey G Katrukha
Sep 25, 2017·Clinical Chemistry·Ivan A KatrukhaAlexey G Katrukha
Aug 30, 2017·Biological Chemistry·Simonas Savickas, Ulrich Auf dem Keller
Feb 15, 2017·Clinical Biochemistry·Vincent FridénOla Hammarsten
Jul 31, 2021·Journal of Immunological Methods·Hui MaRichard O'Kennedy

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biomarkers for Cardiovascular Risk Assessment

Sensitive and accurate biomarkers used in cardiovascular risk prediction can potentially be used to manage the risk of cardiovascular disease. Discover the latest research on Biomarkers for Cardiovascular Risk Assessment here. Discover the latest research on Biomarkers for Cardiovascular Risk Assessment here.

Related Papers

Medicine and Science in Sports and Exercise
Robert ShavePaul O Collinson
Journal of the Medical Association of Thailand = Chotmaihet Thangphaet
Apichard Sukonthasarn, Sumeth Ponglopisit
© 2022 Meta ULC. All rights reserved