Singlet oxygen-induced protein aggregation: Lysozyme crosslink formation and nLC-MS/MS characterization Emerson Finco Marques, Marisa H.G. Medeiros, Paolo Di Mascio Journal of Mass Spectrometry, 2019 Singlet molecular oxygen (1 O2 ) has been associated with a number of physiological processes. Despite the recognized importance of 1 O2 -mediated protein modifications, little is known about the role of this oxidant in cross-link formation and protein aggregation. Thus, using lysozyme as a model, the present study, sought to investigate the involvement of 1 O2 in cross-link formation. Lysozyme was photochemically oxidized in the presence of Rose Bengal or chemically oxidized using [18 O]-labeled 1 O2 released from thermolabile endoperoxides. It was concluded that both 1 O2 generating systems induce lysozyme crosslinking and aggregation. Using SDS-PAGE and nano-scale liquid chromatography coupled to electrospray ionization mass spectrometry the results clearly demonstrated that 1 O2 is directly involved in the formation of covalent cross-links involving the amino acids Histidine, lysine and Tryptophan.
Lysozyme oxidation by singlet molecular oxygen: Peptide characterization using [18O]-labeling oxygen and nLC-MS/MS Emerson Finco Marques, Marisa H.G. Medeiros, Paolo Di Mascio Journal of Mass Spectrometry, 2017 Singlet molecular oxygen (1 O2 ) is generated in biological systems and reacts with different biomolecules. Proteins are a major target for 1 O2 , and His, Tyr, Met, Cys, and Trp are oxidized at physiological pH. In the present study, the modification of lysozyme protein by 1 O2 was investigated using mass spectrometry approaches. The experimental findings showed methionine, histidine, and tryptophan oxidation. The experiments were achieved using [18 O]-labeled 1 O2 released from thermolabile endoperoxides in association with nano-scale liquid chromatography coupled to electrospray ionization mass spectrometry. The structural characterization by nLC-MS/MS of the amino acids in the tryptic peptides of the proteins showed addition of [18 O]-labeling atoms in different amino acids.
The carbonylation and covalent dimerization of human superoxide dismutase 1 caused by its bicarbonate-dependent peroxidase activity is inhibited by the radical scavenger tempol Raphael F. Queiroz, Verônica Paviani, Fernando R. Coelho, Emerson F. Marques, Paolo Di Mascio, Ohara Augusto Biochemical Journal, 2013 Tempol (4-hydroxy-2,2,6,6-tetramethyl piperidine-1-oxyl) reduces tissue injury in animal models of various diseases via mechanisms that are not completely understood. Recently, we reported that high doses of tempol moderately increased survival in a rat model of ALS (amyotrophic lateral sclerosis) while decreasing the levels of oxidized hSOD1 (human Cu,Zn-superoxide dismutase) in spinal cord tissues. To better understand such a protective effect in vivo, we studied the effects of tempol on hSOD1 oxidation in vitro. The chosen oxidizing system was the bicarbonate-dependent peroxidase activity of hSOD1 that consumes H2O2 to produce carbonate radical, which oxidizes the enzyme. Most of the experiments were performed with 30 μM hSOD1, 25 mM bicarbonate, 1 mM H2O2, 0.1 mM DTPA (diethylenetriaminepenta-acetic acid) and 50 mM phosphate buffer at a final pH of 7.4. The results showed that tempol (5–75 μM) does not inhibit hSOD1 turnover, but decreases its resulting oxidation to carbonylated and covalently dimerized forms. Tempol acted by scavenging the carbonate radical produced and by recombining with hSOD1-derived radicals. As a result, tempol was consumed nearly stoichiometrically with hSOD1 monomers. MS analyses of turned-over hSOD1 and of a related peptide oxidized by the carbonate radical indicated the formation of a relatively unstable adduct between tempol and hSOD1-Trp32•. Tempol consumption by the bicarbonate-dependent peroxidase activity of hSOD1 may be one of the reasons why high doses of tempol were required to afford protection in an ALS rat model. Overall, the results of the present study confirm that tempol can protect against protein oxidation and the ensuing consequences.
RECENT SCHOLAR PUBLICATIONS
Natural Products as Novel Opportunities for Cathepsin Inhibitors EF Marques Natural Products in Drug Discovery, 311-353 , 2026 2026
Reactive Oxygen Species (ROS) Involvement in the Oxidation and Aggregation of Proteins in the eyes EF Marques, P Di Mascio Free Radical Biology & Medicine 208, S122 res. 265 , 2023 2023
Oxidation of apoptosis-inducing factor (AIF) to disulfide-linked conjugates SM Lo, PA Martinez, EF Marques, S Miyamoto, G Valdameri, VR Moure, ... Archives of Biochemistry and Biophysics 692, 108515 , 2020 2020 Citations: 7
Singlet oxygen‐induced protein aggregation: Lysozyme crosslink formation and nLC‐MS/MS characterization EF Marques, MHG Medeiros, P Di Mascio Journal of Mass Spectrometry 54 (11), 894-905 , 2019 2019 Citations: 11
EVALUATION OF THE POTENTIAL ACARICIDAL OF THE HEXANIC EXTRACT OF Trichilia pallida SW.(SAPINDALES: MELIACEAE) LEAVES AND ESSENTIAL OIL OF Sesamum indicum LINN.(LAMIALES … C de Oliveira Inácio, E de Freitas Fernandes-Pires, LC de Castro Jr, ... SaBios-Revista de Saúde e Biologia 14 (1), 1-10 , 2019 2019
EVALUATION OF THE POTENTIAL ACARICIDAL OF THE HEXANIC EXTRACT OF Trichilia pallida SW.(SAPINDALES: MELIACEAE) LEAVES AND ESSENTIAL OIL OF Sesamum indicum LINN.(LAMIALES … NB Amaro, LC de Castro Junior, C de Oliveira Inácio, R Nunes, ... environment 9, 10 , 2019 2019
Lysozyme oxidation by singlet molecular oxygen: Peptide characterization using [ 18 O]‐labeling oxygen and nLC‐MS/MS EF Marques, MHG Medeiros, P Di Mascio Journal of Mass Spectrometry 52 (11), 739-751 , 2017 2017 Citations: 14
Oxygen contribution for oxidation and protein aggregation EF Marques, MHG Medeiros, P Di Mascio Proceedings , 2017 2017
Investigação de produtos de reação do oxigênio singlete em proteínas por espectrometria de massas e marcação isotópica EF Marques 2017
Acridone alkaloids as inhibitors of cathepsin L and V. EF Marques, PC Vieira, RP Severino 2016 Citations: 4
[18O]-labeled singlet molecular oxygen: Chemical generation and trapping as a tool for mechanistic studies S Miyamoto, GR Martinez, GE Ronsein, EF Marques, FM Prado, KR Prieto, ... 2016 Citations: 2
ALCALOIDES ACRIDÔNICOS INIBEM CATEPSINA LEV EF Marques, PC Vieira, RP Severino Química Nova 39 (1), 58-62 , 2016 2016 Citations: 3
Production of lysozyme and lysozyme-superoxide dismutase dimers bound by a ditryptophan cross-link in carbonate radical-treated lysozyme V Paviani, RF Queiroz, EF Marques, P Di Mascio, O Augusto Free Radical Biology and Medicine 89, 72-82 , 2015 2015 Citations: 58
Lysozyme Oxidation by Singlet Molecular Oxygen: Cross-Linking Formation and Proposed Mechanism Using 18-O Labeled Oxygen and Mass Spectrometry Measurements EF Marques, AC Scalfo, MHG de Medeiros, P Di Mascio Free Radical Biology and Medicine 1 (87), S149-S150 , 2015 2015
The Carbonylation and Covalent Dimerization of hSOD1 Caused by Its Bicarbonate-Dependent Peroxidase Activity Is Inhibited by Tempol: 201 RF Queiroz, V Paviani, FR Coelho, EF Marques, P Di Mascio, O Augusto Free Radical Biology and Medicine 65, S93 , 2013 2013
The carbonylation and covalent dimerization of human superoxide dismutase 1 caused by its bicarbonate-dependent peroxidase activity is inhibited by the radical scavenger tempol RF Queiroz, V Paviani, FR Coelho, EF Marques, P Di Mascio, O Augusto Biochemical Journal 455 (1), 37-46 , 2013 2013 Citations: 23
Characterization of a ditryptophan cross-link in hen lysozyme oxidized by the carbonate radical V Paviani, RF Queiroz, EF Marques, P Di Mascio, O Augusto Abstracts , 2013 2013
Evaluation of synthetic acridones and 4-quinolinones as potent inhibitors of cathepsins L and V EF Marques, MA Bueno, PD Duarte, LRSP Silva, AM Martinelli, ... European journal of medicinal chemistry 54, 10-21 , 2012 2012 Citations: 50
Acridone alkaloids as potent inhibitors of cathepsin V RP Severino, RVC Guido, EF Marques, D Brömme, MFGF da Silva, ... Bioorganic & medicinal chemistry 19 (4), 1477-1481 , 2011 2011 Citations: 50
Estudo de produtos naturais e derivados sintéticos buscando inibidores seletivos das catepsinas L e V EF Marques Universidade Federal de São Carlos , 2011 2011 Citations: 2
MOST CITED SCHOLAR PUBLICATIONS
An extremely facile synthesis of furans, pyrroles, and thiophenes by the dehydrative cyclization of propargyl alcohols A Aponick, CY Li, J Malinge, EF Marques Organic Letters 11 (20), 4624-4627 , 2009 2009 Citations: 274
Production of lysozyme and lysozyme-superoxide dismutase dimers bound by a ditryptophan cross-link in carbonate radical-treated lysozyme V Paviani, RF Queiroz, EF Marques, P Di Mascio, O Augusto Free Radical Biology and Medicine 89, 72-82 , 2015 2015 Citations: 58
Evaluation of synthetic acridones and 4-quinolinones as potent inhibitors of cathepsins L and V EF Marques, MA Bueno, PD Duarte, LRSP Silva, AM Martinelli, ... European journal of medicinal chemistry 54, 10-21 , 2012 2012 Citations: 50
Acridone alkaloids as potent inhibitors of cathepsin V RP Severino, RVC Guido, EF Marques, D Brömme, MFGF da Silva, ... Bioorganic & medicinal chemistry 19 (4), 1477-1481 , 2011 2011 Citations: 50
Solution phase synthesis of a combinatorial library of chalcones and flavones as potent cathepsin V inhibitors J Alvim Jr, RP Severino, EF Marques, AM Martinelli, PC Vieira, ... Journal of Combinatorial Chemistry 12 (5), 687-695 , 2010 2010 Citations: 42
The carbonylation and covalent dimerization of human superoxide dismutase 1 caused by its bicarbonate-dependent peroxidase activity is inhibited by the radical scavenger tempol RF Queiroz, V Paviani, FR Coelho, EF Marques, P Di Mascio, O Augusto Biochemical Journal 455 (1), 37-46 , 2013 2013 Citations: 23
Lysozyme oxidation by singlet molecular oxygen: Peptide characterization using [ 18 O]‐labeling oxygen and nLC‐MS/MS EF Marques, MHG Medeiros, P Di Mascio Journal of Mass Spectrometry 52 (11), 739-751 , 2017 2017 Citations: 14
Singlet oxygen‐induced protein aggregation: Lysozyme crosslink formation and nLC‐MS/MS characterization EF Marques, MHG Medeiros, P Di Mascio Journal of Mass Spectrometry 54 (11), 894-905 , 2019 2019 Citations: 11
Oxidation of apoptosis-inducing factor (AIF) to disulfide-linked conjugates SM Lo, PA Martinez, EF Marques, S Miyamoto, G Valdameri, VR Moure, ... Archives of Biochemistry and Biophysics 692, 108515 , 2020 2020 Citations: 7
Acridone alkaloids as inhibitors of cathepsin L and V. EF Marques, PC Vieira, RP Severino 2016 Citations: 4
das GF da; Fernandes, JB; Adricopulo, AD; Vieira, PC RP Severino, RVC Guido, EF Marques, D Brömme, MF Silva Bioorg. Med. Chem 19, 1477 , 2011 2011 Citations: 4
ALCALOIDES ACRIDÔNICOS INIBEM CATEPSINA LEV EF Marques, PC Vieira, RP Severino Química Nova 39 (1), 58-62 , 2016 2016 Citations: 3
[18O]-labeled singlet molecular oxygen: Chemical generation and trapping as a tool for mechanistic studies S Miyamoto, GR Martinez, GE Ronsein, EF Marques, FM Prado, KR Prieto, ... 2016 Citations: 2
Estudo de produtos naturais e derivados sintéticos buscando inibidores seletivos das catepsinas L e V EF Marques Universidade Federal de São Carlos , 2011 2011 Citations: 2
São Carlos-SP EF Marques Universidade Federal de São Carlos , 2011 2011 Citations: 2
Natural Products as Novel Opportunities for Cathepsin Inhibitors EF Marques Natural Products in Drug Discovery, 311-353 , 2026 2026
Reactive Oxygen Species (ROS) Involvement in the Oxidation and Aggregation of Proteins in the eyes EF Marques, P Di Mascio Free Radical Biology & Medicine 208, S122 res. 265 , 2023 2023
EVALUATION OF THE POTENTIAL ACARICIDAL OF THE HEXANIC EXTRACT OF Trichilia pallida SW.(SAPINDALES: MELIACEAE) LEAVES AND ESSENTIAL OIL OF Sesamum indicum LINN.(LAMIALES … C de Oliveira Inácio, E de Freitas Fernandes-Pires, LC de Castro Jr, ... SaBios-Revista de Saúde e Biologia 14 (1), 1-10 , 2019 2019
EVALUATION OF THE POTENTIAL ACARICIDAL OF THE HEXANIC EXTRACT OF Trichilia pallida SW.(SAPINDALES: MELIACEAE) LEAVES AND ESSENTIAL OIL OF Sesamum indicum LINN.(LAMIALES … NB Amaro, LC de Castro Junior, C de Oliveira Inácio, R Nunes, ... environment 9, 10 , 2019 2019
Oxygen contribution for oxidation and protein aggregation EF Marques, MHG Medeiros, P Di Mascio Proceedings , 2017 2017
