Novel agents for the in-situ detection of cysteine oxidation states

Shibani Ratnayake, Shibani Ratnayake, Christopher Dunston, Eric Lattmann, Helen Griffiths

Research output: Contribution to journalMeeting abstract

Abstract

Cysteine (Cys-SH) is an amino acid, which due to the many stable oxidation states of the sulfur containing thiol moiety, readily undergoes oxidation by reactive oxygen species (ROS) to form sulfenic (Cys-OH), sulfinic (Cys-SO2H) and sulfonic (Cys-SO3H) acids. Thiol modifications of cysteine have been implicated as modulators of cellular processes and represent significant biological modifications that occur during oxidative stress and cell signaling. However, the different oxidation states are difficult to monitor in a physiological setting due to the limited availability of experimental tools. Therefore it is of interest to synthesize a range of chemical probes that selectively recognize specific oxidation states of sulfur and also are cell permeable to allow observation of their formation in real time. The current study is aimed at investigating a synthetic approach for novel fluorescent probe synthesis, for the specific detection of cysteine sulfenic acids. The probe will consist of a dimedone like molecule, 3, 5-diketohexahydrobenzoic which is uniquely reactive with Cys-SOH, a linker and a fluorescent tag which will enable detection by fluorescence spectroscopy. To achieve this, 2-aminoanthracene, a fluorescent molecule has been coupled with chloroacetyl chloride which acts as a bifunctional linker to form N-(anthracen-2-yl)-2-chloroacetamide. This compound has been used to synthesise 2-amino-N-(anthracen-2-yl)acetamide via an amide synthesis. Successful synthesis of these compounds has been confirmed by nuclear magnetic resonance and mass spectrometric analysis. Present work involves the attachment of 3,5-diketohexahydrobenzoic acid synthesise to 2-amino-N-(anthracen-2-yl)acetamide. We will use this for investigation of cysteine oxidation in T cells under oxidative stress. [1-3] Phillips D.C, Dias H.K, Kitas G.D, Griffiths H.R.(2010) Antioxid Redox Signal,12(6):743-85 Grant M.M,Griffiths H.R. (2007) Environmental Pharmacology and Toxicology 23, 335-339 Phillips D.C., Woollard K.J. and Griffiths H.R. (2003) Brit. J. Pharmacol. 138, 501-511.
Original languageEnglish
Article number337
Pages (from-to)S137-S138
Number of pages2
JournalFree Radical Biology and Medicine
Volume53
Issue numberSupplement 2
DOIs
Publication statusPublished - 1 Nov 2012
Event19th Annual Meeting of the Society for Free Radical Biology and Medicine - San Diego, CA, United States
Duration: 14 Nov 201218 Nov 2012

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Cysteine
Sulfur
Sulfhydryl Compounds
Oxidation
Oxidative Stress
Oxidative stress
Ecotoxicology
Acids
Fluorescence Spectrometry
Fluorescent Dyes
Amides
Oxidation-Reduction
Reactive Oxygen Species
Cell signaling
Magnetic Resonance Spectroscopy
Observation
Molecules
Pharmacology
T-cells
T-Lymphocytes

Cite this

Ratnayake, S., Ratnayake, S., Dunston, C., Lattmann, E., & Griffiths, H. (2012). Novel agents for the in-situ detection of cysteine oxidation states. Free Radical Biology and Medicine, 53(Supplement 2), S137-S138. [337]. https://doi.org/10.1016/j.freeradbiomed.2012.10.375
Ratnayake, Shibani ; Ratnayake, Shibani ; Dunston, Christopher ; Lattmann, Eric ; Griffiths, Helen. / Novel agents for the in-situ detection of cysteine oxidation states. In: Free Radical Biology and Medicine. 2012 ; Vol. 53, No. Supplement 2. pp. S137-S138.
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abstract = "Cysteine (Cys-SH) is an amino acid, which due to the many stable oxidation states of the sulfur containing thiol moiety, readily undergoes oxidation by reactive oxygen species (ROS) to form sulfenic (Cys-OH), sulfinic (Cys-SO2H) and sulfonic (Cys-SO3H) acids. Thiol modifications of cysteine have been implicated as modulators of cellular processes and represent significant biological modifications that occur during oxidative stress and cell signaling. However, the different oxidation states are difficult to monitor in a physiological setting due to the limited availability of experimental tools. Therefore it is of interest to synthesize a range of chemical probes that selectively recognize specific oxidation states of sulfur and also are cell permeable to allow observation of their formation in real time. The current study is aimed at investigating a synthetic approach for novel fluorescent probe synthesis, for the specific detection of cysteine sulfenic acids. The probe will consist of a dimedone like molecule, 3, 5-diketohexahydrobenzoic which is uniquely reactive with Cys-SOH, a linker and a fluorescent tag which will enable detection by fluorescence spectroscopy. To achieve this, 2-aminoanthracene, a fluorescent molecule has been coupled with chloroacetyl chloride which acts as a bifunctional linker to form N-(anthracen-2-yl)-2-chloroacetamide. This compound has been used to synthesise 2-amino-N-(anthracen-2-yl)acetamide via an amide synthesis. Successful synthesis of these compounds has been confirmed by nuclear magnetic resonance and mass spectrometric analysis. Present work involves the attachment of 3,5-diketohexahydrobenzoic acid synthesise to 2-amino-N-(anthracen-2-yl)acetamide. We will use this for investigation of cysteine oxidation in T cells under oxidative stress. [1-3] Phillips D.C, Dias H.K, Kitas G.D, Griffiths H.R.(2010) Antioxid Redox Signal,12(6):743-85 Grant M.M,Griffiths H.R. (2007) Environmental Pharmacology and Toxicology 23, 335-339 Phillips D.C., Woollard K.J. and Griffiths H.R. (2003) Brit. J. Pharmacol. 138, 501-511.",
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Ratnayake, S, Ratnayake, S, Dunston, C, Lattmann, E & Griffiths, H 2012, 'Novel agents for the in-situ detection of cysteine oxidation states', Free Radical Biology and Medicine, vol. 53, no. Supplement 2, 337, pp. S137-S138. https://doi.org/10.1016/j.freeradbiomed.2012.10.375

Novel agents for the in-situ detection of cysteine oxidation states. / Ratnayake, Shibani; Ratnayake, Shibani; Dunston, Christopher; Lattmann, Eric; Griffiths, Helen.

In: Free Radical Biology and Medicine, Vol. 53, No. Supplement 2, 337, 01.11.2012, p. S137-S138.

Research output: Contribution to journalMeeting abstract

TY - JOUR

T1 - Novel agents for the in-situ detection of cysteine oxidation states

AU - Ratnayake, Shibani

AU - Ratnayake, Shibani

AU - Dunston, Christopher

AU - Lattmann, Eric

AU - Griffiths, Helen

PY - 2012/11/1

Y1 - 2012/11/1

N2 - Cysteine (Cys-SH) is an amino acid, which due to the many stable oxidation states of the sulfur containing thiol moiety, readily undergoes oxidation by reactive oxygen species (ROS) to form sulfenic (Cys-OH), sulfinic (Cys-SO2H) and sulfonic (Cys-SO3H) acids. Thiol modifications of cysteine have been implicated as modulators of cellular processes and represent significant biological modifications that occur during oxidative stress and cell signaling. However, the different oxidation states are difficult to monitor in a physiological setting due to the limited availability of experimental tools. Therefore it is of interest to synthesize a range of chemical probes that selectively recognize specific oxidation states of sulfur and also are cell permeable to allow observation of their formation in real time. The current study is aimed at investigating a synthetic approach for novel fluorescent probe synthesis, for the specific detection of cysteine sulfenic acids. The probe will consist of a dimedone like molecule, 3, 5-diketohexahydrobenzoic which is uniquely reactive with Cys-SOH, a linker and a fluorescent tag which will enable detection by fluorescence spectroscopy. To achieve this, 2-aminoanthracene, a fluorescent molecule has been coupled with chloroacetyl chloride which acts as a bifunctional linker to form N-(anthracen-2-yl)-2-chloroacetamide. This compound has been used to synthesise 2-amino-N-(anthracen-2-yl)acetamide via an amide synthesis. Successful synthesis of these compounds has been confirmed by nuclear magnetic resonance and mass spectrometric analysis. Present work involves the attachment of 3,5-diketohexahydrobenzoic acid synthesise to 2-amino-N-(anthracen-2-yl)acetamide. We will use this for investigation of cysteine oxidation in T cells under oxidative stress. [1-3] Phillips D.C, Dias H.K, Kitas G.D, Griffiths H.R.(2010) Antioxid Redox Signal,12(6):743-85 Grant M.M,Griffiths H.R. (2007) Environmental Pharmacology and Toxicology 23, 335-339 Phillips D.C., Woollard K.J. and Griffiths H.R. (2003) Brit. J. Pharmacol. 138, 501-511.

AB - Cysteine (Cys-SH) is an amino acid, which due to the many stable oxidation states of the sulfur containing thiol moiety, readily undergoes oxidation by reactive oxygen species (ROS) to form sulfenic (Cys-OH), sulfinic (Cys-SO2H) and sulfonic (Cys-SO3H) acids. Thiol modifications of cysteine have been implicated as modulators of cellular processes and represent significant biological modifications that occur during oxidative stress and cell signaling. However, the different oxidation states are difficult to monitor in a physiological setting due to the limited availability of experimental tools. Therefore it is of interest to synthesize a range of chemical probes that selectively recognize specific oxidation states of sulfur and also are cell permeable to allow observation of their formation in real time. The current study is aimed at investigating a synthetic approach for novel fluorescent probe synthesis, for the specific detection of cysteine sulfenic acids. The probe will consist of a dimedone like molecule, 3, 5-diketohexahydrobenzoic which is uniquely reactive with Cys-SOH, a linker and a fluorescent tag which will enable detection by fluorescence spectroscopy. To achieve this, 2-aminoanthracene, a fluorescent molecule has been coupled with chloroacetyl chloride which acts as a bifunctional linker to form N-(anthracen-2-yl)-2-chloroacetamide. This compound has been used to synthesise 2-amino-N-(anthracen-2-yl)acetamide via an amide synthesis. Successful synthesis of these compounds has been confirmed by nuclear magnetic resonance and mass spectrometric analysis. Present work involves the attachment of 3,5-diketohexahydrobenzoic acid synthesise to 2-amino-N-(anthracen-2-yl)acetamide. We will use this for investigation of cysteine oxidation in T cells under oxidative stress. [1-3] Phillips D.C, Dias H.K, Kitas G.D, Griffiths H.R.(2010) Antioxid Redox Signal,12(6):743-85 Grant M.M,Griffiths H.R. (2007) Environmental Pharmacology and Toxicology 23, 335-339 Phillips D.C., Woollard K.J. and Griffiths H.R. (2003) Brit. J. Pharmacol. 138, 501-511.

U2 - 10.1016/j.freeradbiomed.2012.10.375

DO - 10.1016/j.freeradbiomed.2012.10.375

M3 - Meeting abstract

VL - 53

SP - S137-S138

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - Supplement 2

M1 - 337

ER -

Ratnayake S, Ratnayake S, Dunston C, Lattmann E, Griffiths H. Novel agents for the in-situ detection of cysteine oxidation states. Free Radical Biology and Medicine. 2012 Nov 1;53(Supplement 2):S137-S138. 337. https://doi.org/10.1016/j.freeradbiomed.2012.10.375