Original Article

The Preventive Effect of L-Lysine on Lysozyme Glycation in Type 2 Diabetes


Lysozyme is a bactericidal enzyme whose structure and functions change in diabetes. Chemical chaperones are small molecules including polyamines (e.g. spermine), amino acids (e.g. L-lysine) and polyols (e.g. glycerol). They can improve protein conformation in several stressful conditions such as glycation. In this study, the authors aimed to observe the effect of L-lysine as a chemical chaperone on structure and function of glycated lysozyme. In this study, in vitro and in vivo effects of L-lysine on lysozyme glycation were investigated. Lysozyme was incubated with glucose and/or L-lysine, followed by an investigation of its structure by electrophoresis, fluorescence spectroscopy, and circular dichroism spectroscopy and also assessment of its bactericidal activity against M. lysodeikticus. In the clinical trial, patients with type 2 diabetes mellitus (T2DM) were randomly divided into two groups of 25 (test and control). All patients received metformin and glibenclamide for a three months period. The test group was supplemented with 3 g/day of L-lysine. The quantity and activity of lysozyme and other parameters were then measured. Among the test group, L-lysine was found to reduce the advanced glycation end products (AGEs) in the sera of patients with T2DM and in vitro condition. This chemical chaperone reversed the alteration in lysozyme structure and function due to glycation and resulted in increased lysozyme activity. Structure and function of glycated lysozyme are significantly improved by l-lysine; therefore it can be considered an effective therapeutic supplementation in T2DM, decreasing the risk of infection in these patients.

Tan KC, Chow WS, Ai VH, et al. Advanced glycation end products and endothelial dysfunction in type 2 diabetes. Diabetes Care 2002;25(6):1055-9.

Sensi M, Bruno MR, Valente L, et al. Retinol-binding protein: a short half life determinant of protein nonenzymatic glycation in diabetes. Diabetes Res 1990;13(4):195-8.

Rahbar S, Yerneni KK, Scott S, et al. Novel inhibitors of,advanced glycation end product (part I). Mol Cell Biol Res Commun 2000;3(6):360-6.

Jafarnejad A, Bathaie SZ, Nakhjavani M, et al. The improvement effect of L-Lys as a chemical chaperone on STZ-induced diabetic rats, protein structure and function. Diabetes Metab Res Rev 2008;24(1):64-73.

Jafarnejad A, Bathaie SZ, Nakhjavani M, et al. Investigation of the mechanisms involved in the high-dose and long-term acetylsalicylic acid therapy of type I diabetic rats. J Pharmacol Exp Ther 2008;324(2):850-7.

Rahmanpour R, Bathaie SZ. Histone H1 structural changes and its interaction with DNA in the presence of high glucose concentration In vivo and In vitro. J Biomol Struct Dyn 2011;28(4):575-86.

Bathaie SZ, Nobakht BB, Mirmiranpour H, et al. Effect of chemical chaperones on glucose-induced lysozyme modifications. Protein J 2011;30(7):480-9.

Peppa M, Uribarri J, Vlassara H. Glucose, advanced glycation end products, and diabetes complications. Clin Diabetes 2003;21(4):186-7.

Lapolla A, Traldi P, Fedele D. Importance of measuring products of non-enzymatic glycation of proteins. Clin Biochem 2005;38(2):103-15.

Yanagisawa K, Makitam Z, Shiroshita K, et al. Specific fluorescence assay for advanced glycatio n end products in blood and urine of diabetic patients. Metabolism 1998;47(11):1348-53.

Agardh CD, Stenram U, Torffvit O, et al. Effects ofinhibition of glycation and oxidative stress on the development of diabetic nephropathy in rats. J Diabetes Complicat 2002;16(6):395-400.

Jafarnejad A, Bathaie SZ, Nakhjavani M, et al. Effect of spermine on lipid profile and HDL functionality in the streptozotocin-induced diabetic rat model. Life Sci 2008;82(5-6):301-7.

Gugliucci A, Menini T. The polyamines spermine and spermidine protect proteins from structural and functional damage by AGE precursors: a new role for old molecules? Life Sci 2003;72(23):2603-16.

Perlmutter DH. Chemical chaperones: a pharmacological strategy for disorders of protein folding and trafficking. Pediatr Res 2002;52(6):832-6.

Shirali S, Bathaie SZ, Nakhjavani N. Effects of saffron aqueous extract on some biochemical factors of serum instreptozotocin-induced diabetic rats. Iran J Med Aromatic Plants 2012;28(2):293-308.

Yeboah FK, Alli I, Yaylayan VA, et al. Effect of limited solid-state glycation on the conformation of lysozyme by ESI-MSMS peptide mapping and molecular modeling. Bioconjug Chem 2004;15(1):27-34.

Masuda T, Ide N, Kitabatake N. Effects of chemical modification of lysine residues on the sweetness of lysozyme. Chem Senses 2005;30(3):253-64.

Ibrahim HR, Aoki T, Pillegrini A. Strategies for new antimicrobial proteins and peptides: Lysozyme and aprotinin as model molecules. Curr Pharm Des 2002;8(9):671-93.

Peeters TL, Depraetere YR, Vantrappen GR. Radioimmunoassay for urinary lysozyme in human serum from leukemic patients. Clin Chem 1978;24(12):2155-7.

Frare E, Polverino De Laureto P, Zurdo J, et al. A highly amyloidogenic region of hen lysozyme. J Mol Biol 2004;340(5):1153-65.

Powroznik B, Gharbi M, Dandrifosse G, et al.Enhancement of lysozyme stability and activity bypolyamines. Biochime 2004;86(9-10):651-6.

Saito A, Sako Y, Usui M, et al. Functional properties of glycosylated lysozyme secreted in Pichia pastoris. Biosci Biotechnol Biochem 2003;67(11):2334-43.

Longo MA, Combes D. A novel chemoenzymatic glycosylation strategy: application to lysozyme modification. FEBS Lett 1995;37(1-2):63-6.

Li YM. Glycation ligand binding motif in lactoferrin. Implications in diabetic infection. Adv Exp Med Biol 1998;443:57-63.

Reisfeld R, Lewis U, Williams D. Disk electrophoresis of basic proteins and peptides on polyacrylamide gel. Nature 1962;195(4838):281-3.

Kalousova M, Skrha J, Zima T. Advanced glycation endproducts and advanced oxidation protein products in patients with diabetes mellitus. Physiol Res 2002;51(6):597-604.

Goldberg M, Rudolph R, Jaenicke R. A kinetic study of the competition between renaturation and aggregation during the refolding of denatured-reduced egg white lysozyme. Biochemistry 1991;30(11):2790-7.

American Diabetes Association. Screening for diabetes (position statement). Diabetes Care 2001;24(Suppl 1):S21-4.

Spinozzi F, Velardi A, Rambotti P, et al. Circulating immune complexes and serum lysozyme levels in untreated Hodgkin's disease. Their relationship to immune function. J Clin Lab Immunol 1983;12(2):87-92.

Vittal BG, Praveen G, Deepak P. A study of body mass index in healthy individuals and its relationship with fasting blood sugar. J Clin Diag Res 2010;4(6):3421-4.

Zheng F, Cai W, Mitsuhashi T. Lysozyme enhances renal excretion of advanced glycation end products in vivo andsuppresses adverse AGE-mediated cellular effects in vitro: a potential AGE sequestration therapy for diabetic nephropathy. Mol Med 2001;7(3):737-47.

Monnier VM. Intervention against the Maillard reaction in vivo. Arch Biochem Biophys 2003;419(1):1-15.

Mirmiranpour H, Bathaie SZ, Khaghani S. Investigation of the mechanism(s) involved in decreasing increased fibrinogen 2 activity in hyperglycemic conditions using Llysine supplementation. Thrombosis Res 2012;130(3):e13-9.

Masuda T, Ide N, Kitabatake N. Structure–sweetness relationship in egg white lysozyme: role of lysine and arginine residues on the elicitation of lysozyme sweetness. Chem Senses 2005;30(8):667-81.

GhoshMoulick R, Bhattacharya J. Compensatory secondary structure alterations in protein glycation. Biochim Biophys Acta (BBA) 2007;1774(2):233-42.

Binazzi M, Boncio L, Marconi P, et al. Serum and skin lysozyme activity in non-diabetic and diabetic subjects. Arch Dermatol Res 1978;262(3):239-45.

Zemliakova ZM, Kravets EB. Activity of nonspecific immunological factors and serum immunoglobulin levels in children with diabetes mellitas. Probl Endokrinol (Mosk) 1981;27(6):6-9.

Lechowski R, Lenarcik M, Degòrski A, et al. Serum lysozyme activity and Nitroblue tetrazolium reduction test in dogs with diabetes mellitus. Zentralbl Veterinarmed A 1991;38(7):530-3.

Joo NS, Lee DJ, Winges KM, et al. Regulation of antiprotease and antimicrobial protein secretion by airway submucosal gland serous cells. J Biol Chem 2004;279(37):38854-60.

Naglaa ZHE, Hesham AM, Hosny AF, et al. Impact of metformin on immunity and male fertility in rabbits with alloxan- induced diabetes. J Am Sci 2010;6(11):417-26.

Pang G, Clancy R, Cong M, et al. Influenza virus inhibits lysozyme secretion by sputum neutrophils in subjects with chronic bronchial sepsis. Am J Respir Crit Care Med 2000;161(3 Pt 1):718-22.

Griffith RS, Walsh DE, Myrmel KH, et al. Success of Llysine therapy in frequently recurrent herpes simplex infection. Treatment and prophylaxis. Dermatologica 1987;175(4):183-90.

IssueVol 54, No 1 (2016) QRcode
SectionOriginal Article(s)
Lysozyme Chemical chaperone L-lysine Type 2 diabetes

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Mirmiranpour H, khaghani shahnaz, Bathaie SZ, Nakhjavani M, Kebriaeezadeh A, Ebadi M, Gerayesh-Nejad S, Zangooei M. The Preventive Effect of L-Lysine on Lysozyme Glycation in Type 2 Diabetes. Acta Med Iran. 2016;54(1):24-31.