[Frontiers In Bioscience, Landmark, 24, 1024-1036, March 1, 2019]

Characterization of structural requirement for binding of gigantol and aldose reductase

Yong Yang 1,2, Qiaohong Yang1, Juan Yu1, Wencheng Wan1, Xiaoyong Wei1,2

1School of Basic Medical Sciences,Guangzhou University of Chinese Medicine, Guangzhou Guangdong,510006, PR China, 2School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang 310000, PR China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Material, reagents and instruments
3.2. Animals and animal care
3.3. Effect of gigantol on lens opacity in rats
3.4. Extraction of AR from the rat lens and measurement of the inhibitory activity of gigantol on AR
3.5. Molecular dynamics simulation and molecular docking
3.6. Construction of pET28a vector expressing AR and its mutants
3.7. Expression and purification of AR and the mutants
3.8. Determination of catalytic activity of recombinant AR and its mutants
3.9. Detection of non-covalent bonding of gigantol to AR with cold-spray ionization mass spectrometry (CSI-MS)
3.10. Statistical methods
4. Results
4.1. Protection of gigantol against STZ-induced lens opacity in rats
4.2. Gigantol inhibits AR activity
4.3. Prediction of the binding sites between gigantol and AR by MD
4.4. The interaction trajectory of gigantol and AR
4.5. Identification of the binding sites between gigantol and AR through site-directed mutagenesis
4.6. Determination of non-covalent bonding between gigantol and AR by CSI-MS
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

We previously reported that gigantol extracted from Caulis Dendrobii has significant therapeutic benefits for the treatment of galactosemic cataracts through its ability to inhibit aldose reductase (AR) activity. In this study, we identified the binding sites and structurally characterized the interaction between gigantol and AR, to understand the mechanism (s) of the effects of gigantol on cataracts. Gigantol was found to be protective against diabetic cataracts (DC) in rats induced by streptozotocin. Molecular docking predicted the binding sites between AR and gigantol to be residues Trp111, His110, Tyr48 and Trp20. Mutation of each of these residues led to a significant reduction in AR activity. Cold-spray ionization mass spectrometry measurements showed that the binding of gigantol to AR is concentration-dependent and that the maximum stoichiometric ratio of non-covalent bonding is 1:24.4. pH and temperature did not influence the interaction. Taken together, we provide further mechanistic evidence of the beneficial effects of gigantol on DC.

7. REFERENCES

1. S. Huang, Y. Zheng, P. J. Foster, W. Huang and M. He: Prevalence and causes of visual impairment in Chinese adults in Urban Southern China. Arch Ophthalmol,127(10),1362-1367(2009)
DOI: 10.1001/archophthalmol.2009. 138 3

2. A. S. Bessa, A. M. Ragab, R. A. Nassra, D. P. Piñero and M. S. Shaheen: Expression levels of aldose reductase enzyme, vascular endothelial growth factor, and intercellular adhesion molecule-1 in the anterior lens capsule of diabetic cataract patients. J Cataract Refract Surg,44(12),1431-1435(2018)
DOI: 10.1016/j.jcrs.2018.07.054

3. I. Dedov, O. Maslova, Y. Suntsov, L. Bolotskaia, T. Milenkaia and L. Besmertnaia: Prevalence of diabetic retinopathy and cataract in adult patients with type 1 and type 2 diabetes in Russia. Rev Diabet Stud,6,124-129(2009)
DOI: 10.1900/RDS.2009.6.124

4. S.Wu, N.Tong, LPan, X.H Jiang, Y.A. Li, M.L Guo and H.H. Li: Retrospective Analyses of Potential Risk Factors for Posterior Capsule Opacification after Cataract Surgery. J Ophthalmol. 9089285 (2018)..
DOI: 10.1155/2018/9089285

5. C. V. Ton and T.H.C.Tran:Incidence of posterior capsular opacification requiring Nd:YAG capsulotomy after cataract surgery and implantation of enVista® MX60 IOL. J Fr Ophtalmol.41(10),899-903(2018)
DOI: 10.1016/j.jfo.2018.04.011

6. I. M. Wormstone: Posterior capsule opacification: a cell biological perspective. Exp Eye Res,74(3),337-347 (2002)
DOI: 10.1006/exer.2001.1153

7. F. Tian, L. Dong, Y. Zhou, Y. Shao, W. Li, H. Zhang and F. Wang: Rapamycin-Induced apoptosis in HGF-stimulated lens epithelial cells by AKT/mTOR, ERK and JAK2/STAT3 pathways. Int J Mol Sci,15(8),13833-13848(2014)
DOI: 10.3390/ijms150813833

8. A. B. Reddy, R. Tammali, R. Mishra, S. Srivastava, S. K. Srivastava and K. V. Ramana: A1dose reductase deficiency protects sugar-induced lens opacification in rats. Chem Biol Interact,191,346-350(2011)
DOI: 10.1016/j.cbi.2011.02.028

9. A. Gul, M. A. Rahman and S. N. Hasnain: Role of fructose concentration on cataractogenesis in senile diabetic and non-diabetic patients. Graefes Arch Clin Exp Ophthalmol,47,809-814(2009)
DOI: 10.1007/s00417-008-1027-9

10. D.Ganesh, K Torigoe, M.Kumano-Kuramochi, S.Machida and T. Kobori:Microplate-Based Assay for Screening of Advanced Glycation End Products Binding to Its Receptor. Anal Sci, (2019)
DOI: 10.2116/analsci.18C021

11. G. J. Zablocki, P. A. Ruzycki, M. A. Overturf, S. Palla, G. B. Reddy and J. M. Petrash: Aldose reductase-meduated induction of epithelium-to-mesenchymal transition (EMT) in lens. Chem Biol Interact,191,351-356(2011)
DOI: 10.1016/j.cbi.2011.02.005

12. S.K.Srivastava, U.C.Yadav, A.B.Reddy, A.Saxena, R.Tammali, M.Shoeb, N.H.Ansari, A.Bhatnagar, M.JPetrash, S.Srivastava and K.V.Ramana:Aldose reductase inhibition suppresses oxidative stress-induced inflammatory disorders. Chem Biol Interact,191(1-3):330-8(2011).
DOI: 10.1016/j.cbi.2011.02.023.

13. C. J. Bulpitt, Y. Li, P. F. Bulpitt and J. Wang: The use of orchids in Chinese medicine. J R Soc Med,100,558-563(2007)
DOI: 10.1177/0141076807100012014

14. W.P.Zheng, Y.P.Tang, F.Zhi and Lou FC.:Dihydroayapin, a new coumarin compound from Dendrobium densiflorum. J Asian Nat Prod Res,2(4):301-304.(2000)
DOI: 10.1080/10286020008041369

15. J. M. Hu, J. J. Chen, H. Yu, Y. X. Zhao and J. Zhou: Five new compounds from Dendrobium longicornu. Planta Med,74,535-9(2008)
DOI: 10.1055/s-2008-1074492

16. N.Petpiroon, B.Sritularak and Chanvorachote P: Phoyunnanin E inhibits migration of non-small cell lung cancer cells via suppression of epithelial-to-mesenchymal transition and integrin αv and integrin β3. BMC Complement Altern Med., 17 (1):553. (2017)
DOI: 10.1186/s12906-017-2059-7

17. P. Klongkumnuankarn, K. Busaranon, P.Chanvorachote, B.Sritularak, V.Jongbunprasert and K.Likhitwitayawuid: Cytotoxic and Antimigratory Activities of Phenolic Compounds from Dendrobium brymerianum.Evid Based Complement Alternat Med, 350410 (2015)
DOI: 10.1155/2015/350410

18. L.Qian, G.Ding, Q.Zhou, Z.Feng, X.Ding, S.Gu, Y.Wang, X.Li and B.Chu: Molecular authentication of Dendrobium loddigesii Rolfe by Amplification Refractory Mutation System (ARMS). Planta Med, 74 (4):470-3 (2008).
DOI: 10.1055/s-2008-1034360

19. M. Déciga-Campos, J. F. Palacios-Espinosa, A. Reyes-Ramírez and R. Mata: Antinociceptive and anti-inflammatory effects of compounds isolated from Scaphyglottis livida and Maxillaria densa. J Ethnopharmacol, 114,161-8 (2007)
DOI: 10.1016/j.jep.2007.07.021

20. S. Estrada-Soto, R. Mata, J. J. López-Guerrero, R. Villalobos-Molina and R. Mata: Endothelium-independent relaxation of aorta rings by two stilbenoids from the orchids Scaphyglottis livida. Fitoterapia,77, 236-239(2006)
DOI: 10.1016/j.fitote.2006.02.006

21. S.Charoenrungruang, P.Chanvorachote, B.Sritularak and V. Pongrakhananon: Gigantol, a bibenzyl from Dendrobium draconis, inhibits the migratory behavior of non-small cell lung cancer cells. J Nat Prod, 77 (6):1359-66. (2014).
DOI: 10.1021/np500015v

22. H. Fang, X. Hu, M. Wang, W. Wan, Q. Yang, X. Sun, Q. Gu, X. Gao, Z. Wang, L. Gu, C. Y. Oliver Chen and X. Wei: Anti-osmotic and antioxidant activities of gigantol from Dendrobium aurantiacum var. denneanum against cataractogenesis in galactosemic rats. J Ethnopharmacol,172,238-246(2015)
DOI: 10.1016/j.jep.2015.06.034

23. P. Suryanarayana, M. Saraswat, T. Mrudula, T. P. Krishna, K. Krishnaswamy and G. B. Reddy: Curcumin and turmeric delay streptozotocin-induced diabetic cataract in rats. Invest Ophthalmol Vis Sci,46(6),2092-9(2005).
DOI: 10.1167/iovs.04-1304

24. S.B.Kim, S.H.Hwang, Z.Wang, J.M.Yu and S.S.Lim: Rapid Identification and Isolation of Inhibitors of Rat Lens Aldose Reductase and Antioxidant in Maackia amurensis. Biomed Res Int, 4941825 (2017).
DOI: 10.1155/2017/4941825

25. T.Y.Song,N.C.Yang, C.L.Chen and T.L.V.Thi: Protective Effects and Possible Mechanisms of Ergothioneine and Hispidin against Methylglyoxal-Induced Injuries in Rat Pheochromocytoma Cells. Oxid Med Cell Longev. 2017, 1-10 (2017).
DOI: 10.1155/2017/4824371

26. S. S. Lim, Y. J. Jung, S. K. Hyun, Y. S. Lee and J. S. Choi: Rat lens aldose reductase inhibitory constituents of Nelumbo nucifera stamens. Phytother Res, 20,825-830(2006)
DOI: 10.1002/ptr.1847

27. H. M. Li, S. H. Hwang, B. G. Kang, J. S. Hong and S. S. Lim: Inhibitory effects of Colocasia esculenta(L.) Schott constituents on aldose reductase. Molecules, 19,13212-13224(2014)
DOI: 10.3390/molecules190913212

28. A.Podjarny, R.E.Cachau, T.Schneider, M. Z.Van and A..Joachimiak: Subatomic and atomic crystallographic studies of aldose reductase: implications for inhibitor binding.Cell Mol Life Sci,61(7-8):763-73(2004)
DOI: 10.1007/s00018-003-3404-1

29. A.Hassanin, Y. Kaminishi and T.Itakura : Characterization of Tilapia (Oreochromis niloticus) aldehyde reductase (AKR1A1) gene, promoter and expression pattern in benzo-a-pyrene exposed fish. Toxicol Mech Methods.27(1):36-44(2017).
DOI: 10.1080/15376516.2016.1238529

30. L. Ji, L.Cheng and Z.Yang: Diosgenin, a Novel Aldose Reductase Inhibitor, Attenuates the Galactosemic Cataract in Rats. J Diabetes Res,7309816. (2017).
DOI: 10.1155/2017/7309816

31. H. Steuber, M. Zentgraf, C. Gerlach, C. A. Sotriffer, A. Heine and G. Klebe: Expect the unexpected or caveat for drug designers: multiple structure determinations using aldose reductase crystals treated under varying soaking and co-crystallisation conditions. J Mol Biol, 363,174-87 (2006)
DOI: 10.1016/j.jmb.2006.08.011

32. R. Maccari, R. Ottanà, R. Ciurleo, D. Rakowitz, B. Matuszczak, C. Laggner and T. Langer: Synthesis, induced-fit docking investigations, and in vitro aldose reductase inhibitory activity of non-carboxylic acid containing 2,4-thiazolidinedione derivatives. Bioorg Med Chem, 16, 5840-52 (2008).
DOI: 10.1016/j.bmc.2008.04.072

33. Z. Wang, B. Ling, R. Zhang, Y. Suo, Y. Liu, Z. Yu and C.liu: Docking and molecular dynamics studies toward the binding of new natural phenolic marine inhibitors and aldose reductase. J Mol Graph Model, 28,162-9 (2009).
DOI: 10.1016/j.jmgm.2009.06.003

34. C. Koukoulitsa, F. Bailly, K. Pegklidou, V. J. Demopoulos and P. Cotelle: Evaluation of aldose reductase inhibition and docking studies of 6’-nitro and 6’,6’’-dinitrorosmarinic acids. Eur J Med Chem, 45, 1663-6 (2010) .
DOI: 10.1016/j.ejmech.2009.12.007

35. K.Sundaram, S.Endo, T.Matsunaga, N.Tanaka, A.Hara and El-Kabbani O: Structure of the His269Arg mutant of the rat aldose reductase-like protein AKR1B14 complexed with NADPH. Acta Crystallogr Sect F Struct Biol Cryst Commun., 68 (Pt 4):400-3. (2012).
DOI: 10.1107/S1744309112008810

36. X. Wei, D. Chen, Y. Yi, H. Qi, X. Gao, H. Fang, Q. Gu, L. Wang and L. Gu: Syringic Acid Extracted from Herba dendrobii Prevents Diabetic Cataract Pathogenesis by Inhibiting Aldose Reductase Activity. Evid Based Complement Alternat Med,426537(2012)
DOI: 10.1155/2012/426537

37. R. Maccari, R. Ciurleo, M. Giglio, M. Cappiello, R. Moschini, A. D. Corso, U. Mura and R. Ottanà: Identification of new non-carboxylic acid containing inhibitors of aldose reductase. Bioorg Med Chem, 18, 4049-4055 (2010).
DOI: 10.1016/j.bmc.2010.04.016

38. Z.Hashim and S.Zarina: Osmotic stress induced oxidative damage: possible mechanism of cataract formation in diabetes. J Diabetes Complications.26 (4):275-9 (2012).
DOI: 10.1016/j.jdiacomp.2012.04.005

39. Y. Li, T. H. Huang and J. Yamahara: Salacia root, a unique Ayurvedic medicine, meets multiple targets in diabetes and obesity. Life Sci,82,1045-1049(2008)
DOI: 10.1016/j.lfs.2008.03.005

40. G. K. Balendiran and B. Rajkumar: Fibrates inhibit aldose reductase activity in the forward and reverse reactions. Biochem Pharmacol, 70,1653-1663(2005)
DOI: 10.1016/j.bcp.2005.06.029

41. M. Stefek, Prnova M. Soltesova, M. Majekova, C. Rechlin, A. Heine and G. Klebe: Identification of novel aldose reductase inhibitors based on carboxymethylated mercaptotriazinoindole scaffold. J Med Chem, 58 (6), 2649-2657 (2015).
DOI: 10.1021/jm5015814

42. M. P. Kumar, V. Sankeshi, R. R. Naik, P. Thirupathi, B. Das and T. N. Raju: The inhibitory effect of Isoflavones isolated from Caesalpinia pulcherrima on aldose reductase in STZ induced diabetic rats. Chem Biol Interact, 237, 18-24 (2015) .
DOI: 10.1016/j.cbi.2015.05.010

43. S. R. Sharma and N. Sharma: Epalrestat, an aldose reductase inhibitor, in diabetic neuropathy: an Indian perspective. Ann Indian Acad Neurol, 11 (4), 231-235 (2008).
DOI: 10.4103/0972-2327.44558

44. S.Senthilkumari, R.Sharmila, G.Chidambaranathan, A.Vanniarajan: Epalrestat, an Aldose Reductase Inhibitor Prevents Glucose-Induced Toxicity in Human Retinal Pigment Epithelial Cells In Vitro. J Ocul Pharmacol Ther, 33 (1):34-41 (2017).
DOI: 10.1089/jop.2016.0103.

45. C. Veeresham, Rao A. Rama and K. Asres: Aldose reductase inhibitors of plant origin. Phytother Res,28 (3),317-333 (2014).
DOI: 10.1002/ptr.5000

46. H. N. Yoon, M. Y. Lee, J. K. Kim, H. W. Suh and S. S. Lim: Aldose reductase inhibitory compounds from Xanthium strumarium. Arch Pharm Res, 36 (9), 1090-1095 (2013)
DOI: 10.1007/s12272-013-0123-5

47. J. P. Luo, Y. Y. Deng and X. Q. Zha: Mechanism of Polysaccharides from Dendrobium huoshanense on Streptozotocin-Induced Diabetic Cataract. Pharmaceutical Biology, 46,243-249 (2008).
DOI: 10.1016/j.jpba.2007.09.024

48. H.An, Q.Zhu, W.Pei, J.Fan, Y.Liang, Y.Cui, N.Lv and Wang W: Whole Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo. PLoS One., 11(11):e0163478 (2016).
DOI: 10.1371/journal.pone.0163478

49. P.P.Singh, S.K.Aithagani, M.Yadav, V.P.Singh and R.A.Vishwakarma: Iron-catalyzed cross-coupling of electron-deficient heterocycles and quinone with organoboron species via innate C-H functionalization: application in total synthesis of pyrazine alkaloid botryllazine A. J Org Chem.78 (6):2639-48. (2013).
DOI: 10.1021/jo302797r

50. A. Ramunno, S. Cosconati, S. Sartini, V. Maglio and S. Angiuoli, V. L. Pietra, S. D. Maro, M. Giustiniano , C..L. Motta, F.D. Settimo, LMarinelli and E.Novellino: Progresses in the pursuit of aldose reductase inhibitors: the structure-based lead optimization step. Eur J Med Chem, 51,216-226 (2012).
DOI: 10.1016/j.ejmech.2012.02.045

Abbreviations: diabetic cataracts (DC), aldose reductase (AR), aldose reductase inhibitors (ARI), AR,Ala substitution at Trp20 (W20A), AR,Ala substitution at Tyr48 (Y48A), AR,Ala substitution at His110 (H110A), AR,Ala substitution at Trp111 (W111A), phosphate buffer saline (PBS), dimethyl sulfoxide (DMSO), molecular docking (MD), protein data bank (PDB), 50% inhibition of enzyme activity (IC50), transferable intermolecular potential 3 points (TIP3P), wild-type (WT), root mean square deviation (RMSD), cold spray ionization mass spetrometry (CSI-MS), Streptozotocin (STZ), Kanamycin (Kan), Dalton (Da), Nanosecond (ns), reduced nicotinamide adenine dinucleotide phosphate (NADPH), three times a day (TID), complementary DNA (cDNA), polymerase chain reaction (PCR), basic local alignment search tool (BLAST), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Michaelis-Menten constant (Km), maximum velocity (Vax)

Key Words: Binding Sites, Gigantol, Aldose Reductase, Diabetic Cataract, Site-Directed Mutagenesis, Cold-Spray Ionization Mass Spectrometry

Send correspondence to: Xiaoyong Wei, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine; School of Medicine, Hangzhou Normal University. Tel: 8618520498313, Fax: 862039358588, E-mail:jidewowxy@163.com