Amino Acids

, Volume 47, Issue 12, pp 2551–2560 | Cite as

Tyrosine- and tryptophan-coated gold nanoparticles inhibit amyloid aggregation of insulin

  • Kriti Dubey
  • Bibin G. Anand
  • Rahul Badhwar
  • Ganesh Bagler
  • P. N. Navya
  • Hemant Kumar Daima
  • Karunakar Kar
Original Article


Here, we have strategically synthesized stable gold (AuNPsTyr, AuNPsTrp) and silver (AgNPsTyr) nanoparticles which are surface functionalized with either tyrosine or tryptophan residues and have examined their potential to inhibit amyloid aggregation of insulin. Inhibition of both spontaneous and seed-induced aggregation of insulin was observed in the presence of AuNPsTyr, AgNPsTyr, and AuNPsTrp nanoparticles. These nanoparticles also triggered the disassembly of insulin amyloid fibrils. Surface functionalization of amino acids appears to be important for the inhibition effect since isolated tryptophan and tyrosine molecules did not prevent insulin aggregation. Bioinformatics analysis predicts involvement of tyrosine in H-bonding interactions mediated by its C=O, –NH2, and aromatic moiety. These results offer significant opportunities for developing nanoparticle-based therapeutics against diseases related to protein aggregation.


Amyloid aggregation Tryptophan Tyrosine Insulin Gold nanoparticles 



Circular dichroism


Tyrosine-coated gold nanoparticles


Tryptophan-coated gold nanoparticles


Tyrosine-coated silver nanoparticles


Transition temperature



We thank IIT Jodhpur for research facilities. We thank IIT Bombay for use of the Cryo HR-TEM Central Facility. This work was supported by BRNS grant (KK) (Grant No.37(1)/14/38/2014-BRNS) and Seed Grant from Indian Institute of Technology Jodhpur (KK and GB). HKD gratefully acknowledges Department of Science and Technology (DST), Government of India for ITS Grant (Grant No. SB/ITS-Y/0988/2014-15).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

726_2015_2046_MOESM1_ESM.pdf (1.7 mb)
Supplementary material 1 (PDF 1702 kb)


  1. Aguzzi A, O’Connor T (2010) Protein aggregation diseases: pathogenicity and therapeutic perspectives. Nat Rev Drug Discovery 9:237–248CrossRefPubMedGoogle Scholar
  2. Alvarez YD, Fauerbach JA, Pellegrotti JV, Jovin TM, Jares-Erijman EA, Stefani FD (2013) Influence of gold nanoparticles on the kinetics of α-synuclein aggregation. Nano Lett 13:6156–6163CrossRefPubMedGoogle Scholar
  3. Bhattacharyya R, Chakrabarti P (2003) Stereospecific interactions of proline residues in protein structures and complexes. J Mol Biol 331:925–940CrossRefPubMedGoogle Scholar
  4. Bolton EE, Wang Y, Thiessen PA, Bryant SH (2008) PubChem: integrated platform of small molecules and biological activities. Ann Rep Comput Chem 4:217–241CrossRefGoogle Scholar
  5. Brooks BR, Brooks CL III, Mackerell AD Jr, Nilsson L, Petrella RJ, Roux B, Won Y, Archontis G, Bartels C, Boresch S et al (2009) CHARMM: the biomolecular simulation program. J Comput Chem 30:1545–1614PubMedCentralCrossRefPubMedGoogle Scholar
  6. Cai H, Yao P (2014) Gold nanoparticles with different amino acid surfaces: serum albumin adsorption, intracellular uptake and cytotoxicity. Colloids Surf B Biointerfaces 123:900–906CrossRefPubMedGoogle Scholar
  7. Chatani E, Imamura H, Yamamoto N, Kato M (2014) Stepwise organization of the β-structure identifies key regions essential for the propagation and cytotoxicity of insulin amyloid fibrils. J Biol Chem 289:10399–10410PubMedCentralCrossRefPubMedGoogle Scholar
  8. Chiti F, Dobson CM (2006) Protein misfolding, functional amyloid, and human disease. Ann Rev Biochem 75:333–366CrossRefPubMedGoogle Scholar
  9. Chiti F, Dobson CM (2009) Amyloid formation by globular proteins under native conditions. Nat Chem Biol 5:15–22CrossRefPubMedGoogle Scholar
  10. Daima HK, Selvakannan PR, Shukla R, Bhargava SK, Bansal V (2013) Fine-tuning the antimicrobial profile of biocompatible gold nanoparticles by sequential surface functionalization using polyoxometalates and lysine. PLoS ONE 8:e79676PubMedCentralCrossRefPubMedGoogle Scholar
  11. Dubey K, Anand BG, Temgire MK, Kar K (2014) Evidence of rapid coaggregation of globular proteins during amyloid formation. Biochemistry 53:8001–8004CrossRefPubMedGoogle Scholar
  12. Etienne MA, Aucoin JP, Fu Y, McCarley RL, Hammer RP (2006) Stoichiometric inhibition of amyloid beta-protein aggregation with peptides containing alternating alpha, alpha-disubstituted amino acids. J Am Chem Soc 128:3522–3523CrossRefPubMedGoogle Scholar
  13. Fandrich M, Fletcher MA, Dobson CM (2001) Amyloid fibrils from muscle myoglobin. Nature 410:165–166CrossRefPubMedGoogle Scholar
  14. Gao N, Zhang Q, Mu Q, Bai Y, Li L, Zhou H, Butch ER, Powell TB, Snyder SE, Jiang G, Yan B (2011) Steering carbon nanotubes to scavenger receptor recognition by nanotube surface chemistry modification partially alleviates NFkappaB activation and reduces its immunotoxicity. ACS Nano 5:4581–4591PubMedCentralCrossRefPubMedGoogle Scholar
  15. Ghosh R, Sharma S, Chattopadhyay K (2009) Effect of arginine on protein aggregation studied by fluorescence correlation spectroscopy and other biophysical methods. Biochemistry 48:1135–1143CrossRefPubMedGoogle Scholar
  16. Greenwald J, Riek R (2010) Biology of amyloid: structure, function, and regulation. Structure 18:1244–1260CrossRefPubMedGoogle Scholar
  17. Hong DP, Fink AL (2005) Independent heterologous fibrillation of insulin and its B-chain peptide. Biochemistry 44:16701–16709CrossRefPubMedGoogle Scholar
  18. Kar K, Kishore N (2007) Enhancement of thermal stability and inhibition of protein aggregation by osmolytic effect of hydroxyproline. Biopolymers 87:339–351CrossRefPubMedGoogle Scholar
  19. Majzik A, Fulop L, Csapo E, Bogar F, Martinek T, Penke B, Biro G, Dekany I (2010) Functionalization of gold nanoparticles with amino acid, beta-amyloid peptides and fragment. Colloids Surf B Biointerfaces 81:235–241CrossRefPubMedGoogle Scholar
  20. Margiolaki I, Giannopoulou AE, Wright JP, Knight L, Norrman M, Schluckebier G, Fitch AN, Von Dreele RB (2013) High-resolution powder X-ray data reveal the T(6) hexameric form of bovine insulin. Acta Crystallogr D Biol Crystallogr 69:978–990CrossRefPubMedGoogle Scholar
  21. Mark RH, Krebs LA, Morozova-Roche KD, Carol VR, Dobson CM (2004) Observation of sequence specificity in the seeding of protein amyloid fibrils. Protein Sci 13:1933–1938CrossRefGoogle Scholar
  22. Maruyama T, Fujimoto Y, Maekawa T (2014) Synthesis of gold nanoparticles using various amino acids. J Colloid Interface Sci. doi: 10.1016/j.jcis.2014.12.046 PubMedGoogle Scholar
  23. Morozova-Roche LA, Zurdo J, Spencer A, Noppe W, Receveur V, Archer DB, Joniau M, Dobson CM (2000) Amyloid fibril formation and seeding by wild-type human lysozyme and its disease-related mutational variants. J Struct Biol 130:339–351CrossRefPubMedGoogle Scholar
  24. Pechkova E, Bragazzi N, Bozdaganyan M, Belmonte L, Nicolini C (2014) A review of the strategies for obtaining high-quality crystals utilizing nanotechnologies and microgravity. Crit Rev Eukaryot Gene Expr 24:325–339CrossRefPubMedGoogle Scholar
  25. Rajasekhar K, Suresh SN, Manjithaya R, Govindaraju T (2015) Rationally designed peptidomimetic modulators of abeta toxicity in Alzheimer’s disease. Sci Rep 5:8139PubMedCentralCrossRefPubMedGoogle Scholar
  26. Selvakannan P, Ramanathan R, Plowman BJ, Sabri YM, Daima HK, O’Mullane AP, Bansal V, Bhargava SK (2013) Probing the effect of charge transfer enhancement in off resonance mode SERS via conjugation of the probe dye between silver nanoparticles and metal substrates. Phys Chem Chem Phys 15:12920–12929CrossRefPubMedGoogle Scholar
  27. Shiraki K, Kudou M, Fujiwara S, Imanaka T, Takagi M (2002) Biophysical effect of amino acids on the prevention of protein aggregation. J Biochem 132:591–595CrossRefPubMedGoogle Scholar
  28. Siposova K, Kubovcikova M, Bednarikova Z, Koneracka M, Zavisova V, Antosova A, Kopcansky P, Daxnerova Z, Gazova Z (2012) Depolymerization of insulin amyloid fibrils by albumin-modified magnetic fluid. Nanotechnology 23:055101CrossRefPubMedGoogle Scholar
  29. Smith GD, Duax WL, Dodson EJ, Dodson GG, de Graaf RAG, Reynolds CD (1982) The structure of Des-Phe B1 bovine insulin. Acta Crystallogr Sect B Struct Crystallogr Cryst Chem 38:3028–3032CrossRefGoogle Scholar
  30. Smith GD, Pangborn WA, Blessing RH (2005) The structure of T6 bovine insulin. Acta Crystallogr D Biol Crystallogr 61:1476–1482CrossRefPubMedGoogle Scholar
  31. Stefani M, Rigacci S (2013) Protein folding and aggregation into amyloid: the interference by natural phenolic compounds. Int J Mol Sci 14:12411–12457PubMedCentralCrossRefPubMedGoogle Scholar
  32. Swift B (2002) Examination of insulin injection sites: an unexpected finding of localized amyloidosis. Diabet Med J Br Diabet Assoc 19:881–882CrossRefGoogle Scholar
  33. Tartaglia GG, Pawar AP, Campioni S, Dobson CM, Chiti F, Vendruscolo M (2008) Prediction of aggregation-prone regions in structured protein. J Mol Biol 380:425–436CrossRefPubMedGoogle Scholar
  34. Viet MH, Ngo ST, Lam NS, Li MS (2011) Inhibition of aggregation of amyloid peptides by beta-sheet breaker peptides and their binding affinity. J Phys Chem B 115:7433–7446CrossRefPubMedGoogle Scholar
  35. Wu G, Robertson DH, Brooks CL, Vieth M (2003) Detailed analysis of grid-based molecular docking: a case study of CDOCKER-A CHARMm-based MD docking algorithm. J Comput Chem 24:1549–1562CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Kriti Dubey
    • 1
  • Bibin G. Anand
    • 1
  • Rahul Badhwar
    • 1
  • Ganesh Bagler
    • 1
  • P. N. Navya
    • 2
  • Hemant Kumar Daima
    • 2
  • Karunakar Kar
    • 1
  1. 1.Department of BiologyIndian Institute of Technology JodhpurJodhpurIndia
  2. 2.Department of BiotechnologySiddaganga Institute of TechnologyTumkurIndia

Personalised recommendations