Effects of Fenton Reaction on Human Serum Albumin
An In Vitro Study
Keywords:
HSA, Fenton system, Carbonylation assay, SDS-PAGEAbstract
Introduction: Human serum albumin (HSA) is a critical protein in human blood plasma, which can be highly damaged by oxidative stress. The aim of this study was to analyze modifications of this protein after oxidation using a Fenton system.
Methods: In this 2015 experiment, different ratios of Fenton reagent (Fe2+/H2O2) was incubated with one concentration of human serum albumin (1mg/ml). Hence, HSA was incubated 30 min with various combinations of a Fenton system and quantified oxidation products such as carbonyl groups, fragmentations, degradations, and oxidized free thiol group using reliable techniques. Image and data analysis were carried out using ImageJ software and Excel (version 2007), respectively.
Results: An SDS-PAGE profile showed no cross link and aggregation. However, protein band intensity has decreased to 50% in the highest ratio of H2O2/Fe. Carbonylation assay indicated carbonyl/protein (molc/molp) ratio increased linearly in lower ratios and the values plateau at higher levels of H2O2/Fe 2+. The only free sulfhydryl group on HSA was oxidized in all ratios of the Fenton system.
Conclusion: To sum, the structure of HSA has been changed following treatment with Hydroxyl Radical as the main product of Fenton reaction. These data confirm the antioxidant activity of HSA.
References
Seyfizadeh N, Mahjoub S, Zabihi E, Moghadamnia AA, Pouramir M, Mir H, et al. Cytoprotective Effects
of Arbutin Against Tert-Butyl Hydroperoxid Induced Toxicity in Hep-G2 Cell Line. World Applied
Sciences Journal. 2012; 19(2): 163-7.
Gaber MH. Effect of gamma-Irradiation on the Molecular Properties of Bovine Serum Albumin. J Biosci
Bioeng. 2005; 100(2): 203–6. doi: 10.1263/jbb.100.203. PMID: 16198265.
Nguyen AT, Donaldson RP. Metal-catalyzed oxidation induces carbonylation of peroxisomal proteins and
loss of enzymatic activities. Arch Biochem Biophys. 2005; 439(1): 25-31. doi: 10.1016/j.abb.2005.04.018.
PMID: 15922287.
Bochi GV, Torbitz VD, Cargnin LP, de Carvalho JA, Gomes P, Moresco RN. An Alternative Pathway
Through the Fenton Reaction for the Formation of Advanced Oxidation Protein Products, a New Class of
Inflammatory Mediators. Inflammation. 2014; 37(2): 512-21. doi: 10.1007/s10753-013-9765-1. PMID:
Trachootham D, Lu W, Ogasawara MA, Nilsa RD, Huang P. Redox regulation of cell survival. Antioxid
Redox Signal. 2008; 10(8): 1343-74. doi: 10.1089/ars.2007.1957. PMCID: PMC2932530.
Barbehenn R, Dodick T, Poopat U, Spencer B. Fenton-Type Reactions and Iron Concentrations in the
Midgut Fluids of Tree-Feeding Caterpillars. Arch Insect Biochem Physiol. 2005; 60(1): 32–43. doi:
1002/arch.20079. PMID: 16116620.
Kanti Das T, Wati MR, Fatima-Shad K. Oxidative Stress Gated by Fenton and Haber Weiss Reactions and
Its Association With Alzheimer’s Disease. Arch Neurosci. 2015; 2(2): 20078. doi:
5812/archneurosci.20078.
Lenarczyk M, Cohen EP, Fish BL, Irving AA, Sharma M, Driscoll CD, et al. Chronic Oxidative Stress as a
Mechanism for Radiation Nephropathy. Radiat Res. 2009; 171(2): 164-72. doi: 10.1667/RR1454.1. PMID:
, PMCID: PMC2664164.
Guedes S, Vitorino R, Domingues R, Amado F, Domingues P. Oxidation of bovine serum albumin:
identification of oxidation products and structural modifications. Rapid Commun Mass Spectrom. 2009;
(15): 2307-15. doi: 10.1002/rcm.4149. PMID: 19575405.
He F. Laemmli-SDS-PAGE. Bio-protocol Bio. 2011; 101(80)
Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R. Protein carbonyl groups as biomarkers of
oxidative stress. Clin Chim Acta. 2003; 329(1-2): 23-38. doi: 10.1016/S0009-8981(03)00003-2.
Castro-Acosta RM, Rodriguez-Limas WA, Valderrama B, Ramirez OT, Palomares LA. Effect of metal
catalyzed oxidation in recombinant viral protein assemblies. Microb Cell Fact. 2014; 13(1): 25. doi:
1186/1475-2859-13-25. PMID: 24533452, PMCID: PMC3928578.
Headlam HA, Davies MJ. Markers Of Protein xideased Carbonyl Products. Free Radic Biol Med. 2004;
(9): 1175-84. doi: 10.1016/j.freeradbiomed.2004.02.017. PMID: 15082071.
Madian AG, Regnier FE. Proteomic Identification Of Carbonylated Proteins And Their Oxidation Sites. J
Proteome Res. 2010; 9(8): 3766–80. doi: 10.1021/pr1002609. PMID: 20521848, PMCID: PMC3214645.
Turell L, Radi R, Alvarez B. The thiol pool in human plasma: The central contribution of albumin to redox
processes. Free Radic Biol Med. 2013; 65: 244-53. doi: 10.1016/j.freeradbiomed.2013.05.050. PMID:
, PMCID: PMC3909715.
Taverna M, Marie AL, Mira JP, Guidet B. Specific antioxidant properties of human serum albumin. Ann
Intensive Care. 2013; 3(1): 4. doi: 10.1186/2110-5820-3-4. PMID: 23414610, PMCID: PMC3577569.
Kawakami A, Kubota K, Yamada N, Tagami U, Takehana K, Sonaka I, et al. Identification and
characterization of oxidized human serum albumin. A slight structural change impairs its ligand-binding
and antioxidant functions. Febs j. 2006; 273(14): 3346-57. doi: 10.1111/j.1742-4658.2006.05341.x. PMID:
Blouquit Y, Duchambon P, Brun E, Marco S, Rusconi F, Sicard-Roselli C. High sensitivity of human
centrin 2 toward radiolytical oxidation: C-terminal tyrosinyl residue as the main target. Free Radic Biol
Med. 2007; 43(2): 216-28. doi: 10.1016/j.freeradbiomed.2007.04.004. PMID: 17603931.
Roche M, Rondeau P, Singh NR, Tarnus E, Bourdon E. The antioxidant properties of serum albumin. Febs
Letters. 2008; 582(13): 1783-7. doi: 10.1016/j.febslet.2008.04.057. PMID: 18474236.
Yumin Wang, Huayu Xiong, Xiuhua ZhangWang S. Electrochemical study of bovine serum albumin
damage induced by Fenton reaction using tris (2,2-bipyridyl) cobalt (III) perchlorate as the electroactive
indicator. Electrochimica Act. 2012; 67: 147–51. doi: 10.1016/j.electacta.2012.02.010.
Mishra K, Ojha H, Kallepalli S, Alok A, Kumar Chaudhury N. Protective effect of ferulic acid on ionizing
radiation induced damage in bovine serum albumin. Internatuinal Journal of Radiation Research. 2014;
(2): 113-21.
Schuessler H, Schilling K. Oxygen effect in the radiolysis of proteins Part 2 Bovine serum albumin. Int J
Radiat Biol Relat Stud Phys Chem Med. 1984; 45(3): 267-81. doi: 10.1080/09553008414550381. PMID:
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