БИОХИМИЯ, 2022, том 87, вып. 2, с. 160–176

УДК 577.112.7;617.741

Связь между структурой и шаперонной активностью αA‑кристаллина человека после его модификации ассоциированными с сахарным диабетом окислительными агентами и защитная роль антиоксидантных молекул

© 2022 S.S. Moghadam 1, M. Ghahramani 1, K. Khoshaman 1, A. Oryan 2, A.A. Moosavi-Movahedi 3, Б.И. Курганов 4, R. Yousefi 1,3*ryousefi@shirazu.ac.ir; r.yousefi2000@gmail.com

1 Protein Chemistry Laboratory, Department of Biology, College of Sciences Shiraz University, Shiraz, Iran

2 Department of Pathology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

3 Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran

4 Институт биохимии имени А.Н. Баха, ФГУ ФИЦ «Фундаментальные основы биотехнологии» РАН, 119071 Москва, Россия

Поступила в редакцию 12.09.2021
После доработки 16.10.2021
Принята к публикации 16.10.2021

DOI: 10.31857/S0320972522020026

КЛЮЧЕВЫЕ СЛОВА: αA-кристаллин человека, диабетическая катаракта, антиоксиданты хрусталика глаза, структура белка, шаперонная активность.

Аннотация

Целью исследования было оценить влияние пероксинитрита (PON, агент окислительного стресса при диабете), метилглиоксаля (MGO, диабет-ассоциированное реактивное карбонильное соединение) и их одновременного воздействия на структурные и функциональные характеристики αA‑кристаллина человека (αA‑Cry) с помощью различных методов спектроскопии. Кроме того, с помощью метода тензиометрического анализа и метода динамического светорассеяния были определены такие параметры, как поверхностное натяжение и размеры олигомеров обработанного и необработанного белка. Наши результаты показали, что инкубация PON и MGO с αA‑Cry человека приводит к образованию новых хромофоров, вызывает изменения в структурах этого белка (от вторичной и до четвертичной), уменьшение размера белковых олигомеров и значительное усиление шаперонной активности αA‑Cry. Для отмены действия протестированных химических соединений применяли аскорбиновую кислоту и глутатион (основные компоненты антиоксидантной защиты хрусталика глаза). Как ожидалось, оба изученных антиоксиданта в значительной степени предотвращают образование высокомолекулярных агрегатов белка αA‑Cry (по результатам SDS-ПААГ). Полученные нами результаты свидетельствуют о том, что антиоксидантная защита клеток хрусталика, в частности глутатион, может обеспечивать эффективную защиту от быстрого возникновения и прогрессирования диабетической катаракты, предотвращая деструктивные реакции высокореактивных метаболитов, ассоциированных с сахарным диабетом.

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Финансирование

Проведение настоящей работы было поддержано Ученым советом университета Шираза, Научным фондом Ирана (INSF) (проекты 9914455 и 9610387), Национальным институтом медицинских исследований (NIMAD) (проект 964854) и Министерством науки и высшего образования Российской Федерации (Б.И.К).

Конфликт интересов

Авторы заявляют об отсутствии конфликта интересов.

Соблюдение этических норм

Настоящая статья не содержит описания каких-либо исследований с участием людей или животных в качестве объектов.

Список литературы

1. Nowotny, K., Jung, T., Höhn, A., Weber, D., and Grune, T. (2015) Advanced glycation end products and oxidative stress in type 2 diabetes mellitus, Biomolecules, 5, 194-222, doi: 10.3390/biom5010194.

2. Semchyshyn, H. M. (2014) Reactive carbonyl species in vivo: Generation and dual biological effects, Sci. World J., 2014, 1-10, doi: 10.1155/2014/417842.

3. Vander Jagt, D. L. (2008) Methylglyoxal, diabetes mellitus and diabetic complications, Drug Metabol. Drug Interact., 23, 93-124, doi: 10.1515/dmdi.2008.23.1-2.93.

4. Pacher, P., Beckman, J. S., and Liaudet, L. (2007) Nitric oxide and peroxynitrite in health and disease, Physiol. Rev., 87, 315-424, doi: 10.1152/physrev.00029.2006.

5. Sasaki, K., Sasaki, H., Jonasson, F., Kojima, M., and Cheng, H. M. (2004) Racial differences of lens transparency properties with aging and prevalence of age-related cataract applying the WHO classification system, Ophthalmic. Res., 36, 332-340, doi: 10.1159/000081636.

6. Linetsky, M., Shipova, E., Cheng, R., and Ortwerth, B. J. (2008) Glycation by ascorbic acid oxidation products leads to the aggregation of lens proteins, Biochim. Biophys. Acta, 1782, 22-34.

7. Kiziltoprak, H., Tekin, K., Inanc, M., and Sakir Goker, Y. (2019) Cataract in diabetes mellitus, World J. Diabetes, 10, 140-153.

8. Šimunović, M., Paradžik, M., Škrabić, R., Unić, I., Bućan, K., et al. (2018) Cataract as early ocular complication in children and adolescents with type 1 diabetes mellitus, Int. J. Endocrinol., 2018, 1-6, doi: 10.1155/2018/6763586.

9. Benedek, G. B. (1971) Theory of transparency of the eye, Appl. Opt., 10, 459-473, doi: 10.1364/AO.10.000459.

10. Delaye, M., and Tardieu, A. (1983) Short-range order of crystallin proteins accounts for eye lens transparency, Nature, 302, 415-417, doi: 10.1038/302415a0.

11. Bhat, S. P., and Nagineni, C. N. (1989) Alpha B subunit of lens-specific protein alpha crystallin is present in other ocular and non-ocular tissues, Biochem. Biophys. Res. Commun., 158, 319-325, doi: 10.1016/s0006-291x(89)80215-3.

12. Robman, L., and Taylor, H. (2005) External factors in the development of cataract, Eye (Lond), 19, 1074-1082, doi: 10.1038/sj.eye.6701964.

13. Ma, Z., Hanson, S. R., Lampi, K. J., David, L. L., and Smith, J. B. (1998) Age-related changes in human lens crystallins identified by HPLC and mass spectrometry, Exp. Eye Res., 67, 21-30, doi: 10.1006/exer.1998.0482.

14. Fu, L., and Liang, J. J. (2003) Alteration of protein–protein interactions of congenital cataract crystallin mutants, Invest. Ophthalmol. Vis. Sci., 44, 1155-1159, doi: 10.1167/iovs.02-0950.

15. Reddy, V. S., Kumar, C. U., and Reddy, G. B. (2014) Effect of chronic hyperglycemia on crystallin levels in rat lens, Biochem. Biophys. Res. Commun., 446, 602-607, doi: 10.1016/j.bbrc.2014.03.012.

16. Wang, Y., and Ho, C.-T. (2012) Flavour chemistry of methylglyoxal and glyoxal, Chem. Soc. Rev., 41, 4140-4149, doi: 10.1039/c2cs35025d.

17. Alvarez, B., and Radi, R. (2003) Peroxynitrite reactivity with amino acids and proteins, Amino Acids, 25, 295-311, doi: 10.1007/s00726-003-0018-8.

18. Dalle-Donne, I., Rossi, R., Giustarini, D., Milzani, A., and Colombo, R. (2003) Protein carbonyl groups as biomarkers of oxidative stress, Clin. Chim. Acta, 329, 23-38, doi: 10.1016/s0009-8981(03)00003-2.

19. Dayanand, C. D., Pradeep, K. V., and Kutty, A. V. (2012) Protein carbonyl content as a stable oxidative stress marker in type II diabetes, Int. J. Biol. Med. Res., 3, 2362-2365.

20. Ghahramani, M., Yousefi, R., Khoshaman, K., Sasan Moghadam, S., and Kurganov, B. I. (2016) Evaluation of structure, chaperone-like activity and protective ability of peroxynitrite modified human α-crystallin subunits against copper-mediated ascorbic acid oxidation, Int. J. Biol. Macromol., 87, 208-221, doi: 10.1016/j.ijbiomac.2016.02.040.

21. Chang, W., Wang, R., and Wu, L. (2005) Methylglyoxal-induced nitric oxide and peroxynitrite production in vascular smooth muscle cells, Free Radic. Biol. Med., 38, 286-293, doi: 10.1016/j.freeradbiomed.2004.10.034.

22. Moghadam, S., Oryan, A., Kurganov, B. I., Tamaddon, A. M., Alavianehr, M. M., et al. (2017) The structural damages of lens crystallins induced by peroxynitrite and methylglyoxal, two causative players in diabetic complications and preventive role of lens antioxidant components, Int. J. Biol. Macromol., 103, 74-88, doi: 10.1016/j.ijbiomac.2017.04.090.

23. Chen, Y., Mehta, G., and Vasiliou, V. (2014) Antioxidant defenses in the ocular surface, Ocul. Surf., 7, 176-185, doi: 10.1016/s1542-0124(12)70185-4.

24. Merck, K. B., Groenen, P. J., Voorter, C. E., de Haard-Hoekman, W. A., Horwitz, J., et al. (1993) Structural and functional similarities of bovine alpha-crystallin and mouse small heat-shock protein. A family of chaperones, J. Biol. Chem., 268, 1046-1052.

25. Biswas, A., and Das, K. P. (2004) Role of ATP on the interaction of α-crystallin with its substrates and its implications for the molecular chaperone function, J. Biol. Chem., 279, 42648-42657, doi: 10.1074/jbc.M404444200.

26. Nagaraj, R. H., Panda, A. K., Shanthakumar, S., Santhoshkumar, P., Pasupuleti, N., et al. (2012) Hydroimidazolone modification of the conserved Arg12 in small heat shock proteins: studies on the structure and chaperone function using mutant mimics, PLoS One, 7, 1-10, doi: 10.1371/journal.pone.0030257.

27. Robinson, K. M., and Beckman, J. S. (2005) Synthesis of peroxynitrite from nitrite and hydrogen peroxide, Methods Enzymol., 396, 207-214, doi: 10.1016/S0076-6879(05)96019-9.

28. Ghahramani, M., Yousefi, R., Khoshaman, K., and Alavianmehr, M. M. (2015) The impact of calcium ion on structure and aggregation propensity of peroxynitrite-modified lens crystallins: new insights into the pathogenesis of cataract disorders, Colloids Surf. B Biointerfaces, 125, 170-180, doi: 10.1016/j.colsurfb.2014.11.002.

29. Ischiropoulos, H., and Al-Mehdi, A. B. (1995) Peroxynitrite-mediated oxidative protein modifications, FEBS Lett., 364, 279-282, doi: 10.1016/0014-5793(95)00307-u.

30. Alvarez, B., Rubbo, H., Kirk, M., Barnes, S., Freeman, B. A., et al. (1996) Peroxynitrite dependent tryptophan nitration, Chem. Res. Toxicol., 9, 390-396, doi: 10.1021/tx950133b.

31. Kumar, P. A., Kumar, M. S., and Reddy, G. B. (2007) Effect of glycation on alpha-crystallin structure and chaperone-like function, Biochem. J., 408, 251-258, doi: 10.1042/BJ20070989.

32. Al-Hilaly, Y. K., Williams, T. L., Stewart-Parker, M., Ford, L., Skaria, E., et al. (2013) A central role for dityrosine crosslinking of amyloid-β in Alzheimer’s disease, Acta Neuropathol. Commun., 1, 83, doi: 10.1186/2051-5960-1-83.

33. Hospes, M., Hendriks, J., and Hellingwerf, K. J. (2013) Tryptophan fluorescence as a reporter for structural changes in photoactive yellow protein elicited by photo-activation, Photochem. Photobiol. Sci., 12, 479-488, doi: 10.1039/c2pp25222h.

34. Heinecke, J. W., Li, W., Daehnke, H. L. 3rd, and Goldstein, J. A. (1993) Dityrosine, a specific marker of oxidation, is synthesized by the myeloperoxidase-hydrogen peroxide system of human neutrophils and macrophages, J. Biol. Chem., 268, 4069-4077.

35. Paoli, P., Sbrana, F., Tiribilli, B., Caselli, A., Pantera, B., et al. (2010) Protein N-homocysteinylation induces the formation of toxic amyloid-like protofibrils, J. Mol. Biol., 400, 889-907, doi: 10.1016/j.jmb.2010.05.039.

36. Meehan, S., Knowles, T. P., Baldwin, A. J., Smith, J. F., Squires, A. M., et al. (2007) Characterisation of amyloid fibril formation by small heat-shock chaperone proteins human αA-, αB- and R120G αB-crystallins, J. Mol. Biol., 372, 470-484, doi: 10.1016/j.jmb.2007.06.060.

37. Whitmore, L., and Wallace, B. A. (2004) DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data, Nucleic Acids Res., 32, W668-W673, doi: 10.1093/nar/gkh371.

38. Gakamsky, A., Duncan, R. R., Howarth, N. M., Dhillon, B., Buttenschön, K. K., et al. (2017) Tryptophan and non-tryptophan fluorescence of the eye lens proteins provides diagnostics of cataract at the molecular level, Sci. Rep., 7, 40375, doi: 10.1038/srep40375.

39. Kessel, L., Kalinin, S., Nagaraj, R. H., Larsen, M., and Johansson, L. B. (2002) Time-resolved and steady-state fluorescence spectroscopic studies of the human lens with comparison to argpyrimidine, pentosidine and 3-OH-kynurenine, Photochem. Photobiol., 76, 549-554, doi: 10.1562/0031-8655(2002)076<0549:trassf>2.0.co;2.

40. Coussons, P. J., Jacoby, J., Mckay, A., Kelly, S. M., Price, N. C., et al. (1997) Glucose modification of human serum albumin: A structural study, Free Radic. Bio. Med., 22, 1217-1227, doi: 10.1016/s0891-5849(96)00557-6.

41. Munch, G., Keis, R., Wessels, A., Riederer, P., Bahner, U., et al. (1997) Determination of advanced glycation end products in serum by fluorescence spectroscopy and competitive ELISA, Eur. J. Clin. Chem. Clin. Biochem., 35, 669-677, doi: 10.1515/cclm.1997.35.9.669.

42. Cheng, R., Feng, Q., Argirov, O. K., and Ortwerth, B. J. (2005) K2P – a novel cross-link from human lens protein, Ann. N. Y. Acad. Sci., 1043, 184-194, doi: 10.1196/annals.1333.023.

43. Alavi, P., Yousefi, R., Amirghofran, S., Karbalaei-Heidari, H. R., and Moosavi-Movahedi, A. A. (2013) Structural analysis and aggregation propensity of reduced and nonreduced glycated insulin adducts, Appl. Biochem. Biotechnol., 170, 623-638, doi: 10.1007/s12010-013-0207-1.

44. Zhou, C., Qi, W., Lewis, E. N., and Carpenter, J. F. (2015) Concomitant Raman spectroscopy and dynamic light scattering for characterization of therapeutic proteins at high concentrations, Anal. Biochem., 472, 7-20, doi: 10.1016/j.ab.2014.11.016.

45. Bumagina, Z., Gurvits, B., Artemova, N., Muranov, K., and Kurganov, B. I. (2010) Paradoxical acceleration of dithiothreitol-induced aggregation of insulin in the presence of a chaperone, Int. J. Mol. Sci., 11, 4556-4579, doi: 10.3390/ijms11114556.

46. Kurganov, B. I. (2002) Kinetics of protein aggregation. Quantitative estimation of the chaperone-like activity in test-systems based on suppression of protein aggregation, Biochemistry (Moscow), 67, 409-422, doi: 10.1023/a:1015277805345.

47. Biswas, A., and Das, K. P. (2007) Alpha-crystallin assisted refolding of enzyme substrates: Optimization of external parameters, Protein J., 4, 247-255, doi: 10.1007/s10930-006-9066-8.

48. Khoshaman, K., Yousefi, R., Niazi, A., Oryan, A., Moosavi-Movahedi, A. A., et al. (2018) Importance of the positively charged residue at position 54 to the chaperoning function, conformational stability and amyloidogenic nature of human αA-crystallin, J. Biochem., 163, 187-199, doi: 10.1093/jb/mvx071.

49. Jakob, U., Gaestel, M., Engel, K., and Buchner, J. (1993) Small heat shock proteins are molecular chaperones, J. Biol. Chem., 268, 1517-1520.

50. Weber, K., and Osborn, M. (1969) The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis, J. Biol. Chem., 244, 4406-4412.

51. Schägger, H., and von Jagow, G. (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa, Anal. Biochem., 166, 368-379, doi: 10.1016/0003-2697(87)90587-2.

52. Porter, R. R. (1953) Partition chromatography of insulin and other proteins, Biochem. J., 53, 320-328, doi: 10.1042/bj0530320.

53. Pace, C. N., Vajdos, F., Fee, L., Grimsley, G., and Gray, T. (1995) How to measure and predict the molar absorption coefficient of a protein, Protein. Sci., 4, 2411-2423, doi: 10.1002/pro.5560041120.

54. Scientist for Experimental Data Fitting (1995) Microsoft Windows Version 2.0. Salt Lake City: Micro Math, Inc.

55. Moreau, K. L., and King, J. A. (2012) Protein misfolding and aggregation in cataract disease and prospects for prevention, Trends Mol. Med., 18, 273-282, doi: 10.1016/j.molmed.2012.03.005.

56. Argirova, M., and Breipohl, W. (2002) Comparison between modifications of lens proteins resulted from glycation with methylglyoxal, glyoxal, ascorbic acid, and fructose, J. Biochem. Mol. Toxicol., 16, 140-145, doi: 10.1002/jbt.10031.

57. Thiagarajan, G., Lakshmanan, J., Chalasani, M., and Balasubramanian, D. (2004) Peroxynitrite reaction with eye lens proteins: α-crystallin retains its activity despite modification, Biochem. Mol. Biol., 45, 2115-2121, doi: 10.1167/iovs.03-0929.

58. Cubillos-Rojas, M., Schneider, T., Sánchez-Tena, S., Bartrons, R., Ventura, F., et al. (2016) Analysis of protein oligomerization by electrophoresis, Methods Mol. Biol., 1449, 341-348, doi: 10.1007/978-1-4939-3756-1_22.

59. Nowak, P., Zbikowska, H. M., Ponczek, M., Kolodziejczyk, J., and Wachowicz, B. (2007) Different vulnerability of fibrinogen subunits to oxidative/nitrative modifications induced by peroxynitrite: Functional consequences, Thromb. Res., 121, 163-174, doi: 10.1016/j.thromres.2007.03.017.

60. Akhand, A. A., Hossain, K., Cato, M., Miyata, T., Du, J., et al. (2001) Glyoxal and methylglyoxal induce lyoxal and methyglyoxal induce aggergation and inactivation of ERK in human endothelial cells, Free Radic. Biol. Med., 31, 1228-1235, doi: 10.1016/s0891-5849(01)00702-x.

61. Bray, M. R., and Clarke, A. J. (1995) Identification of an essential tyrosyl residue in the binding site of Schizophyllum commune xylanase A, Biochemistry, 34, 2006-2014, doi: 10.1021/bi00006a022.

62. Ye, Y., Quijano, C., Robinson, K. M., Ricart, K. C., Strayer, A. L., et al. (2007) Prevention of peroxynitrite-induced apoptosis of motor neurons and pc12 cells by tyrosine-containing peptides, J. Biol. Chem., 282, 6324-6337, doi: 10.1074/jbc.M610800200.

63. Annibal, A., Colombo, G., Milzani, A., Dalle-Donne, I., Fedorova, M., et al. (2016) Identification of dityrosine cross-linked sites in oxidized human serum albumin, J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 1019, 147-155, doi: 10.1016/j.jchromb.2015.12.022.

64. Maina, M. B., Al-Hilaly, Y. K., Burra, G., Rickard, J., Harrington, C., et al. (2021) Oxidative stress conditions result in trapping of PHF-core tau (297-391) intermediates, Cells, 10, 703, doi: 10.3390/cells10030703.

65. Hawe, A., Sutter, M., and Jiskoot, W. (2008) Extrinsic fluorescent dyes as tools for protein characterization, Pharm. Res., 25, 1487-1499, doi: 10.1007/s11095-007-9516-9.

66. Xue, C., Lin, T. Y., Chang, D., and Guo, Z. (2017) Thioflavin T as an amyloid dye: Fibril quantification, optimal concentration and effect on aggregation, R. Soc. Open. Sci., 4, 1-12, doi: 10.1098/rsos.160696.

67. Greenfield, N. J. (2006) Using circular dichroism spectra to estimate protein secondary structure, Nat. Protoc., 1, 2876-2890, doi: 10.1038/nprot.2006.202.

68. Schor, M., Reid, J. L., MacPhee, C. E., and Stanley-Wall, N. R. (2016) The diverse structures and functions of surfactant proteins, Trends Biochem. Sci., 41, 610-620, doi: 10.1016/j.tibs.2016.04.009.

69. Sattarahmady, N., Moosavi-Movahedi, A. A., and Habibi-Rezaei, M. (2011) A biophysical comparison of human serum albumin to be glycated in vivo and in vitro, J. Med. Biochem., 30, 5-10, doi: 10.2478/v10011-010-0026-7.

70. Yousefi, R., Javadi, S., Amirghofran, S., Oryan, A., and Moosavi-Movahedi, A. A. (2016) Assessment of structure, stability and aggregation of soluble lens proteins and alpha-crystallin upon non-enzymatic glycation: The pathomechanisms underlying cataract development in diabetic patients, Int. J. Biol. Macromol., 82, 328-338, doi: 10.1016/j.ijbiomac.2015.10.036.

71. Ho, M.-C., Peng Y.-J., Chen, S.-J., and Chiou, S.-H. (2010) Senile cataracts and oxidative stress, J. Clin. Gerontol. Geriatr., 1, 17-21, doi: 10.1016/j.jcgg.2010.10.006.

72. Javadi, S., Yousefi, R., Hosseinkhani, S., Tamaddon, A. M., and Uversky, V. N. (2017) Protective effects of carnosine on dehydroascorbate-induced structural alteration and opacity of lens crystallins: Important implications of carnosine pleiotropic functions to combat cataractogenesis, J. Biomol. Struct. Dyn., 35, 1766-1784, doi: 10.1080/07391102.2016.1194230.

73. Das, K. P., and Surewicz, W. K. (1995) On the substrate specificity of α-crystallin as a molecular chaperone, Biochem. J., 311, 367-370, doi: 10.1042/bj3110367.

74. Hook, D. W. A., and Harding, J. J. (1998) Protection of enzymes by α-crystallin acting as a molecular chaperone, Int. J. Biol. Macromol., 22, 295-306, doi: 10.1016/s0141-8130(98)00027-0.

75. Reddy, G. B., Das, K. P., Petrash, J. M., and Surewicz, W. K. (2000) Temperature-dependent chaperone activity and structural properties of human alphaA- and alphaB-crystallins, J. Biol. Chem., 275, 4565-4570, doi: 10.1074/jbc.275.7.4565.

76. Khoshaman, K., Yousefi, R., Tamaddon, A. M., Abolmaali, S. S., Oryan, A., et al. (2017) The impact of different mutations at Arg54 on structure, chaperone-like activity and oligomerization state of human αA-crystallin: The pathomechanism underlying congenital cataract-causing mutations R54L, R54P and R54C, Biochim. Biophys. Acta Proteins Proteom., 1865, 604-618, doi: 10.1016/j.bbapap.2017.02.003.

77. Kisic, B., Miric, D., Zoric, L., Ilic, A., and Dragojevic, I. (2012) Antioxidant capacity of lenses with age-related cataract, Oxid. Med. Cell. Longev., 2012, 467130, doi: 10.1155/2012/467130.

78. Donma, O., Yorulmaz, E. Ö., Pekel, H., and Suyugül, N. (2002) Blood and lens lipid peroxidation and antioxidant status in normal individuals, senile and diabetic cataractous patients, Curr. Eye Res., 25, 9-16, doi: 10.1076/ceyr.25.1.9.9960.

79. Amin, S., Barnett, G., Pathak, J., Roberts, C., and Sarangpani, P. (2014) Protein aggregation, particle formation, characterization & rheology, Curr. Opin. Colloid Interface Sci., 19, 438-449, doi: 10.1016/j.cocis.2014.10.002.

80. Singh, V. P., Bali, A., Singh, N., and Jaggi, A. S. (2014) Advanced glycation end products and diabetic complications, Korean J. Physiol. Pharmacol., 18, 1-14, doi: 10.4196/kjpp.2014.18.1.1.

81. Gul, A., Rahman, M. A., Hasnain, S. N., Salim, A., and Simjee, S. U. (2008) Could oxidative stress associate with age products in cataractogenesis? Curr. Eye Res., 33, 669-675, doi: 10.1080/02713680802250939.

82. Beswick, H. T., and Harding, J. J. (1987) Conformational changes induced in lens alpha- and gamma-crystallins by modification with glucose 6-phosphate. Implications for cataract, Biochem. J., 246, 761-769, doi: 10.1042/bj2460761.

83. Baynes, J. W., and Thorpe, S. R. (1999) Role of oxidative stress in diabetic complications: A new perspective on an old paradigm, Diabetes, 48, 1-9, doi: 10.2337/diabetes.48.1.1.

84. Bizzozero, O. A. (2009) Protein carbonylation in neurodegenerative and demyelinating CNS diseases, Handbook Neurochem. Mol. Neurobiol., 543-562, doi: 10.1007/978-0-387-30375-8_23.

85. Luthra, M., and Balasubramanian, D. (1993) Nonenzymatic glycation alters protein structure and stability. A study of two eye lens crystallins, J. Biol. Chem., 268, 18119-18127.

86. Asmat, U., Abad, K., and Ismail, K. (2015) Diabetes mellitus and oxidative stress-A concise review, Saudi Pharm. J., 24, 547-553, doi: 10.1016/j.jsps.2015.03.013.

87. Bartesaghi, S., and Radi, R. (2018) Fundamentals on the biochemistry of peroxynitrite and protein tyrosine nitration, Redox Biol., 14, 618-625, doi: 10.1016/j.redox.2017.09.009.

88. Nuriel, T., Hansler, A., and Gross, S. S. (2011) Protein nitrotryptophan: formation, significance and identification, J. Proteomics, 74, 2300-2312, doi: 10.1016/j.jprot.2011.05.032.

89. Nagai, R., Unno, Y., Hayashi, M. C., Masuda, S., Hayase, F., et al. (2002) Peroxynitrite induces formation of Nε-(carboxymethyl) lysine by the cleavage of Amadori product and generation of glucosone and glyoxal from glucose, Diabetes, 51, 2833-2839, doi: 10.2337/diabetes.51.9.2833.

90. Kumar, M. S., Mrudula, T., Mitra, N., and Reddy, G. B. (2004) Enhanced degradation and decreased stability of eye lens alpha-crystallin upon methylglyoxal modification, Exp. Eye Res., 79, 577-583, doi: 10.1016/j.exer.2004.07.003.

91. Raman, B., and Rao, C. M. (1994) Chaperone-like activity and quaternary structure of alpha-crystallin, J. Biol. Chem., 269, 27264-27268.

92. Boelens, W. C. (2020) Structural aspects of the human small heat shock proteins related to their functional activities, Cell Stress Chaperones, 25, 581-591, doi: 10.1007/s12192-020-01093-1.

93. Nagaraj, R. H., Nahomi, R. B., Mueller, N. H., Raghavan, C. T., Ammar, D. A., et al. (2016) Therapeutic potential of α-crystallin, Biochim. Biophys. Acta, 1860, 252-257, doi: 10.1016/j.bbagen.2015.03.012.

94. Sharma, K. K., and Santhoshkumar, P. (2009) Lens aging: Effects of crystallins, Biochim. Biophys. Acta, 1790, 1095-1108, doi: 10.1016/j.bbagen.2009.05.008.

95. Cabrera, M. P., and Chihuailaf, R. H. (2011) Antioxidants and the integrity of ocular tissues, Vet. Med. Int., 2011, 905153, doi: 10.4061/2011/905153.