Direct real-time evaluation of nitration with green fluorescent protein in solution and within human cells reveals the impact of nitrogen dioxide vs. peroxynitrite mechanisms

Proceedings of the National Academy of Sciences of the United States of America
Michael G EspeyDavid A Wink

Abstract

3-Nitrotyrosyl adducts in proteins have been detected in a wide range of diseases. The mechanisms by which reactive nitrogen oxide species may impede protein function through nitration were examined by using a unique model system, which exploits a critical tyrosyl residue in the fluorophoric pocket of recombinant green fluorescent protein (GFP). Exposure of purified GFP suspended in phosphate buffer to synthetic peroxynitrite in either 0.5 or 5 microM steps resulted in progressively increased 3-nitrotyrosyl immunoreactivity concomitant with disappearance of intrinsic fluorescence (IC(50) approximately 20 microM). Fluorescence from an equivalent amount of GFP expressed within intact MCF-7 tumor cells was largely resistant to this bolus treatment (IC(50) > 250 microM). The more physiologically relevant conditions of either peroxynitrite infusion (1 microM/min) or de novo formation by simultaneous, equimolar generation of nitric oxide (NO) and superoxide (e.g., 3-morpholinosydnonimine; NONOates plus xanthine oxidase/hypoxanthine, menadione, or mitomycin C) were examined. Despite robust oxidation of dihydrorhodamine under each of these conditions, fluorescence decrease of both purified and intracellular GFP was not evident regardle...Continue Reading

References

Nov 1, 1992·Chemical Research in Toxicology·W H KoppenolJ S Beckman
Nov 1, 1992·Archives of Biochemistry and Biophysics·J S BeckmanM Tsai
Feb 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·J S BeckmanB A Freeman
Jan 1, 1989·Journal of Cardiovascular Pharmacology·M FeelischE Noack
Nov 15, 1985·Archives of Biochemistry and Biophysics·W A PrützE J Land
Dec 20, 1994·Proceedings of the National Academy of Sciences of the United States of America·R HeimR Y Tsien
May 1, 1993·Biochemical Society Transactions·J S Beckman, J P Crow
Jan 1, 1993·Free Radical Research Communications·R E Huie, S Padmaja
Jan 1, 1996·Methods in Enzymology·D ChristodoulouD A Wink
Jul 1, 1996·Chemical Research in Toxicology·J S Beckman
Nov 1, 1996·The American Journal of Physiology·J S Beckman, W H Koppenol
Oct 1, 1996·Nature Biotechnology·F YangG N Phillips
Jul 31, 1998·Archives of Biochemistry and Biophysics·H Ischiropoulos
Oct 9, 1998·The Journal of Biological Chemistry·S Pfeiffer, B Mayer
May 1, 1999·Archives of Biochemistry and Biophysics·D Jourd'heuilD A Wink
Nov 5, 1999·Archives of Biochemistry and Biophysics·J M SouzaH Ischiropoulos
Feb 29, 2000·The Journal of Biological Chemistry·S JayaramanA S Verkman
Feb 7, 2001·The Journal of Biological Chemistry·M G EspeyD A Wink
May 20, 2000·Annals of the New York Academy of Sciences·G C Román
Jun 23, 2000·Annals of the New York Academy of Sciences·M G EspeyD A Wink
Jul 25, 2000·The Journal of Biological Chemistry·C D ReiterJ S Beckman
Feb 22, 2001·Free Radical Biology & Medicine·R RadiA Cayota
Jun 19, 2001·The Journal of Biological Chemistry·M G EspeyD A Wink
Jun 22, 2002·Annals of the New York Academy of Sciences·Michael G EspeyDavid A Wink

❮ Previous
Next ❯

Citations

Sep 4, 2009·Archives of Pharmacal Research·Wilmarie Flores-SantanaDavid A Wink
Jun 4, 2013·Journal of Molecular Graphics & Modelling·Xiao-Hong LiXian-Zhou Zhang
Sep 5, 2002·Free Radical Biology & Medicine·Michael Graham EspeyDavid A Wink
Jul 26, 2003·Antioxidants & Redox Signaling·Douglas D ThomasDavid A Wink
Jul 7, 2005·Antioxidants & Redox Signaling·Mahdieh SadidiDonald M Kuhn
Jan 29, 2008·Antioxidants & Redox Signaling·Hugo P MonteiroLuiz R Travassos
May 27, 2008·American Journal of Physiology. Heart and Circulatory Physiology·Megan L KrajewskiMark T Gladwin
Sep 11, 2010·Journal of Innate Immunity·Yun Yun PangWilliam M Nauseef
Sep 13, 2002·Proceedings of the National Academy of Sciences of the United States of America·Douglas D ThomasDavid A Wink
Aug 15, 2002·Proceedings of the National Academy of Sciences of the United States of America·Michael G EspeyDavid A Wink
May 23, 2012·Free Radical Biology & Medicine·James K Hurst
Apr 29, 2008·Free Radical Biology & Medicine·Douglas D ThomasDavid A Wink
Sep 14, 2007·Immunological Reviews·William M Nauseef
May 23, 2014·Cellular Microbiology·William M Nauseef
Dec 6, 2006·Free Radical Biology & Medicine·Michael Graham Espey
Oct 7, 2006·Biochimica Et Biophysica Acta·Fadi Bou-AbdallahMichael P Lesser
Dec 27, 2005·Biochemical and Biophysical Research Communications·Stefan SchildknechtMarkus Bachschmid
Dec 13, 2005·Free Radical Biology & Medicine·Molly E Beckers-TrappJanet Y Uriu-Adams
Feb 11, 2015·Free Radical Biology & Medicine·Debashree BasudharKatrina M Miranda
Aug 23, 2005·Molecular Aspects of Medicine·Janet Y Uriu-Adams, Carl L Keen
Feb 10, 2009·Free Radical Biology & Medicine·Paul R S BakerBruce A Freeman
Oct 5, 2002·Journal of Lipid Research·Ruba S DeebRita K Upmacis
Sep 10, 2003·Journal of Leukocyte Biology·Brian McElhinneyYvonne M W Janssen-Heininger
May 21, 2008·Chembiochem : a European Journal of Chemical Biology·Christian AmatoreAlaric C W Koh
Dec 20, 2005·American Journal of Physiology. Heart and Circulatory Physiology·Susann PatschanMichael S Goligorsky
Jan 4, 2007·The Journal of Biological Chemistry·Yaozu YeAlvaro G Estévez
Sep 23, 2003·Molecular Pharmacology·Donald M Kuhn, Timothy J Geddes
Jul 20, 2004·Molecular Pharmacology·Johanna T A MeijLei Yu
Dec 7, 2002·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Silvia Pfeiffer, Bernd Mayer
Jul 22, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Jamie SchwartzWilliam M Nauseef
Aug 15, 2019·The Journal of Biological Chemistry·Matías N MöllerBeatriz Alvarez
Jul 7, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Yaoguang ChenNing Shi
Sep 5, 2003·The Journal of Biological Chemistry·Sara GoldsteinAmram Samuni

❮ 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.