Study of Nonenzymatic Glycation of Transferrin and its Effect on Iron -Binding Antioxidant Capacity

Document Type: Original Article

Authors

1 Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

2 Dept of Biochemistry, Medical School, Hamadan University of Medical Sciences, Hamadan, Iran

Abstract

Objective(s)
Nonenzymatic glycosylation (glycation) occurs in many macromolecules in aging and diabetes due to exposure of biomolecules to high level of glucose. Glycation can changes function, activities and structure of many biomolecules. Considering this important role of transferrin (Trf) in iron transport and antioxidant activity in plasma this study was carried out to investigate the effect of glycation in these processes. Materials and Methods
In this study, human apo-Trf (5 mg/ml in sodium phosphate buffer pH= 7.4) was treated with different concentrations of glucose in different period of times (10 days and 20 days). Rate of glycation was measured using thiobarbituric acid method. The effect of glycation on iron binding antioxidant capacity of apo-Trf was investigated using two methods (RBC hemolysis and fluorescent).
Results
Result showed that rate of glycation of apo-Trf was increased with increase in glucose concentration and time of incubation (P< 0.05). Lower iron binding antioxidant capacity was observed for glycted Trf as compared to native Trf (P< 0.05).
Conclusion
Impairment of antioxidant capacity of glycated Trf can suggest a relationship between glycation of Trf and oxidative stress that occurs due to hyperglycemia in diabetic patients.

Keywords


1.Misciagna G, De Michele G, Trevisan M. Non enzymatic glycated proteins in the blood and cardiovascular disease. Curr Pharm Des 2007; 13:3688-3695.

2.Nakajou K, Watanabe H, Kragh-Hansen U, Maruyama T, Otagiri M. The effect of glycation on the structure, function and biological fate of human serum albumin as revealed by recombinant mutants. Biochim Biophys Acta 2003; 1623:88-97.

3.Kennedy DM, Skillen AW, Self CH. Glycation of monoclonal antibodies impairs their ability to bind antigen. Clin Exp Immunol 1994; 98:245-251.

4.Goodarzi MT, Rezaei M, Piry H, Amiry S. Effect of in vitro glycation of human placental collagen (Type IV) on platelet aggregation. Pak J Biolig Sci 2005; 8:1203-1206.

5.Goodarzi MT, Ghahramany S, Mirmoemeni MH. Glycation of human IgG induces structural alterations leading to changes in interaction with anti-IgG. Iran J Immunol 2005; 2:36-42.

6.Goodarzi MT, Ghahramany S, Mirmoemeni MH. In vitro glycation of human IgG and its effect on interaction with anti-IgG. Iran J Allergy Asthma Immunol 2004; 3:181-187.

7.Hemadi M, Kahn PH, Miquel GEl Hage Chahine JM. Transferrin’s mechanism of interaction with receptor 1. Biochemistry 2004; 43:1736-1745.

8.Paez PM, Marta CB, Moreno MB, Soto EF, Pasquini JM. Apotransferrin decreases migration and enhances differentiation of oligodendroglial progenitor cells in an in vitro system. Dev Neurosci 2002; 24:47-58.

9.Gomme PT, McCann KB, Bertolini J. Transferrin: structure, function and potential therapeutic actions. Drug Discov Today 2005; 10:267-273.

10.Van Campenhout A, Van Campenhout C, Lagrou AR, Moorkens G, De Block C, Manuel-y-Keenoy B. Iron-binding antioxidant capacity is impaired in diabetes mellitus. Free Radic Biol Med 2006; 40:1749-1755.

11.Negre-Salvayre A, Coatrieux C, Ingueneau C, Salvayre R. Advanced lipid peroxidation end products in oxidative damage to proteins. Potential role in diseases and therapeutic prospects for the inhibitors. Br J Pharmacol 2008; 153:6-20.

12.Gallou G, Ruelland A, Campion L, Maugendre D, Le Moullec N, Legras B, et al. Increase in thiobarbituric acid-reactive substances and vascular complications in type 2 diabetes mellitus. Diabete Metab 1994: 20:258¬264.

13.Gutteridge JM, Rowley DA, Halliwell B. Superoxide-dependent formation of hydroxyl radicals and lipid peroxidation in the presence of iron salts. Detection of ‘catalytic’ iron and antioxidant activity in extracellular fluids. Biochem J 1982: 206:605-609.

14.Gutteridge JM, Quinlan GJ. Antioxidant protection against organic and inorganic oxygen radicals by normal human plasma: the important primary role for iron-binding and iron-oxidizing proteins. Biochim Biophys Acta 1992; 1159:248-254.

15.Huebschmann AG, Regensteiner JG, Vlassara H, Reusch JEB. Diabetic and advanced glycoxidation end products. Diabetes Care 2006; 29:1420-1432.

16.Martin Gallan' P, Carrascosa A, Gussinye' M, Dominguez C. Biomarkers of diabetes-associated oxidative stress and antioxidant status in young diabetic patients with or without subclinical complications. Free Radic Biol Med 2003;34:1563-1574.

17.Breuer W, Hershko C, Cabantchik ZI. The importance of nontransferrin bound iron in disorders of iron metabolism. Transfus Sci 2000; 23:185-192.

18.Van Campenhout A,Van Campenhout CM, Lagrou AR, Manuel-y-Keenoy B. Transferrin modifications and lipid peroxidation: implications in diabetes mellitus. Free Radic Res 2003; 37:1069-1077.

19.Gallou G, Ruelland A, Campion L, Maugendre D, Le Moullec N, Legras B, et al. Increase in thiobarbituric acid-reactive substances and vascular complications in type 2 diabetes mellitus. Diabete Metab 1994; 20:258¬264.

20.Koga T, Keiko M, Terao J. Protective effet of vitamin E analog, phosphatidylchromanol, against oxidative hemolysis of human erythrocytes. Lipids 1980; 33:589-595.

21.Valkonen M, Kuusi T. Spectrophotometer assay for total peroxyl radical-trapping antioxidant potential in human serum. J Lipid Res 1997; 38:823-833.

22.Sen S, Kar M, Roy A, Chakraborti AS. Effect of nonenzymatic glycation on function and structural properties of hemoglobin. Biophys Chem 2005; 113:289-298.

23.Fujimoto S, Kawakami N, Ohara A. Nonenzymatic glycation of transferri: decrease of iron-binding capacity and increase of oxygen radical production. Biol Pharm Bull 1995; 18:396-400.

24.Van Campenhout A, Van Campenhout C, Lagrou AR, Manuel-y-Keenoy B. Effect of in vitro glycastion on Fe3+ binding and Fe3+ isoforms of transferrin. Clin Chem 2004; 50: 1640-1649.

25.Van Campenhout A, Van Campenhout C, Olyslager YS, Van Damme O, Lagrou AR, Manuel-y-Keenoy B. A novel method to quantify in vivo transferrin glycation: application in diabetes mellitus. Clin Chim Acta 2006; 370:115-123.