Measurement of protein sulfhydryls in response to cellular oxidative stress using gel electrophoresis and multiplexed fluorescent imaging analysis.

Chemical Research in Toxicology
Page C SpiessAlan R Buckpitt

Abstract

The significance of free radicals in biology has been established by numerous investigations spanning a period of over 40 years. Whereas there are many intracellular targets for these radical species, the importance of cysteine thiol posttranslational modification has received considerable attention. The current studies present a highly sensitive method for measurement of the posttranslational modification of protein thiols. This method is based on labeling of proteins with monofunctional maleimide dyes followed by 2D gel electrophoresis to separate proteins and multiplexed fluorescent imaging analysis. The method correctly interrogates the thiol/disulfide ratio present in commercially available proteins. Exposure of pulmonary airway epithelial cells to high concentrations of menadione or t-butyl hydroperoxide resulted in the modification of cysteines in more than 141 proteins of which 60 were subsequently identified by MALDI-TOF/TOF MS. Although some proteins were modified similarly by these two oxidants, several showed detectably different maleimide ratios in response to these two agents. Proteins that were modified by one or both oxidants include those involved in transcription, protein synthesis and folding, and cell death/...Continue Reading

References

Dec 11, 1991·Biochemical Pharmacology·R B BirgeL A Smolin
Dec 1, 1984·Archives of Biochemistry and Biophysics·D Di MonteS Orrenius
Apr 1, 1994·Archives of Biochemistry and Biophysics·Y C ChaiJ A Thomas
Feb 15, 2001·Nature Cell Biology·S R JaffreyS H Snyder
Jan 5, 2002·Physiological Reviews·Wulf Dröge
Aug 24, 2002·Proceedings of the National Academy of Sciences of the United States of America·Albena T Dinkova-KostovaPaul Talalay
Sep 27, 2002·Current Opinion in Biotechnology·Wayne F PattonThomas H Steinberg
Jul 10, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Han-Suk KimAron B Fisher
Jul 23, 2003·Electrophoresis·Kamala TyagarajanJohn E Wiktorowicz
Nov 5, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Asa M WheelockCharles G Plopper
Nov 25, 2003·Analytical Chemistry·Thomas L FareYanqun Wang
Jul 28, 2004·Phytochemistry·François ChevalierMichel Rossignol
Dec 8, 2004·Current Opinion in Structural Biology·David Barford
Mar 19, 2005·Bioinformatics·K K DobbinR M Simon
Apr 23, 2005·American Journal of Respiratory Cell and Molecular Biology·Asa M WheelockAlan R Buckpitt
Jun 14, 2005·Molecular & Cellular Proteomics : MCP·Jiang WuMary Jane Thomassen
Jun 30, 2005·The Journal of Biological Chemistry·Fei HongDaniel C Liebler
Oct 6, 2005·Proteomics·Margaret A IsbellJack Presley
Jan 10, 2006·American Journal of Respiratory Cell and Molecular Biology·Yan WangAron B Fisher
Jan 18, 2006·Chemical Research in Toxicology·Michelle K DennehyDaniel C Liebler
Sep 19, 2006·The International Journal of Biochemistry & Cell Biology·Marian ValkoJoshua Telser
Sep 22, 2006·Antioxidants & Redox Signaling·Janet V Cross, Dennis J Templeton
Sep 30, 2006·Analytical Biochemistry·Lynette K RogersCharles V Smith
Jan 18, 2007·American Journal of Respiratory Cell and Molecular Biology·Michael T Forrester, Jonathan S Stamler
Feb 3, 2007·Current Medicinal Chemistry·V L KinnulaM Myllärniemi
Feb 13, 2007·Free Radical Biology & Medicine·Isabella Dalle-DonneGiancarlo Aldini
Sep 29, 2007·Nature Methods·Alexey I NesvizhskiiRuedi Aebersold
Nov 15, 2007·Antioxidants & Redox Signaling·Dianne M WaltersSteven R Kleeberger
Dec 21, 2007·Antioxidants & Redox Signaling·Isabella Dalle-DonneRanieri Rossi

❮ Previous
Next ❯

Citations

Oct 22, 2009·American Journal of Respiratory Cell and Molecular Biology·Page C SpiessAlan R Buckpitt
Feb 27, 2010·Molecular Nutrition & Food Research·Kishorchandra GohilCarroll E Cross
Oct 3, 2013·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Hwa-Young LeeHyung-Ryong Kim

❮ 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.