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Exploring the anticancer potential of the bacterial protein azurin

1 iBB-Institute for Bioengineering and Biosciences, Biological Sciences Research Group, Av. Rovisco Pais, 1049-001 Lisbon, Portugal
2 Department of Bioengineering, Instituto Superior Técnico, University of Lisbon, Lisbon, Portugal
3 Department of Microbiology & Immunology, University of Illinois College of Medicine, Chicago, IL 60612, USA

Topical Section: Pathogenic microorganisms and human health

Bacterial proteins and their derivative peptides have emerged as promising anticancer agents. Nowadays they represent a valuable set of candidate drugs with different origins and modes of action. Among these, monomeric cupredoxins, which are metalloproteins involved in the electron transport chain of prokaryotes, have been shown to possess potent anticancer activities. In particular, much attention has been focused on azurin produced by the pathogenic bacterium Pseudomonas aeruginosa. More recently, several in vitro and in vivo studies have reported the multi-targeting anticancer properties of azurin. Moreover, p28, a peptide derived from azurin, has completed two phase I clinical trials in cancer patients with promising results. In this updated review, we examine the current knowledge regarding azurin’s modes of action as an anticancer therapeutic protein. We also review the clinical trial results of p28 emphasizing findings that make it suited (alone or in combination) as a therapeutic agent for cancer treatment. Finally we discuss and address the challenges of using the human microbiome to discover novel and unique therapeutic cupredoxin-like proteins.
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Keywords anticancer bacterial proteins; cupredoxins; azurin; p28 peptide; clinical trials; human microbiome

Citation: Arsenio M. Fialho, Nuno Bernardes, Ananda M Chakrabarty. Exploring the anticancer potential of the bacterial protein azurin. AIMS Microbiology, 2016, 2(3): 292-303. doi: 10.3934/microbiol.2016.3.292


  • 1. De Rienzo F, Gabdoulline RR, Menziani MC, et al. (2000) Blue copper proteins: a comparative analysis of their molecular interaction properties. Protein Sci 98: 1439–1454.
  • 2. De Rienzo F, Gabdoulline RR, Wade RC, et al. (2004) Computational approaches to structural and functional analysis of plastocyanin and other blue copper proteins. Cell Mol Life Sci 61: 1123–1142.    
  • 3. Fialho AM, Stevens FJ, Das Gupta TK, et al. (2007) Beyond host-pathogen interactions: microbial defense strategy in the host environment. Curr Opin Biotechnol 18: 279–286.    
  • 4. Stevens FJ (2008) Homology versus analogy: possible evolutionary relationship of immunoglobulins, cupredoxins, and Cu,Zn-superoxide dismutase. J Mol Recognit 21: 20–29.    
  • 5. Warren JJ, Lancaster KM, Richards JH, et al. (2012) Inner- and outer-sphere metal coordination in blue copper proteins. J Inorg Biochem 115: 119–126.    
  • 6. Yanagisawa S, Banfield MJ, Dennison C (2006) The role of hydrogen bonding at the active site of a cupredoxin: the Phe114Pro azurin variant. Biochemistry 45: 8812–8822.    
  • 7. Cannon JG (1989) Conserved lipoproteins of pathogenic neisseria species bearing the H.8 epitope: lipid-modified azurin and H.8 outer membrane protein. Clin Microbiol Rev 2: S1–S4.
  • 8. Hashimoto W, Ochiai A, Hong CS, et al. (2015) Structural studies on Laz, a promiscuous anticancer Neisserial protein. Bioengineered 6: 141–148.    
  • 9. Chaudhari A, Fialho AM, Ratner D, et al. (2006) Azurin, Plasmodium falciparum and HIV/AIDS: inhibition of parasitic and viral growth by azurin. Cell Cycle 5: 1642–1648.    
  • 10. Cruz-Gallardo, Díaz-Moreno, Díaz-Quintana A, et al. (2013) Antimalarial activity of cupredoxins: the interaction of Plasmodium merozoite surface protein 119 (MSP119) and rusticyanin. J Biol Chem 288: 20896–20907.    
  • 11. Naguleswaran A, Fialho AM, Chaudhari A, et al. (2008) Azurin-like protein blocks invasion of Toxoplasma gondii through potential interactions with parasite surface antigen SAG1. Antimicrob Agents Ch 52: 402–408.    
  • 12. Škrlec K, Štrukelj B, Berlec A (2015) Non-immunoglobulin scaffolds: a focus on their targets. Trends Biotechnol 33: 408–418.    
  • 13. Yamada T, Hiraoka Y, Ikehata M, et al. (2004) Apoptosis or growth arrest: modulation of tumor suppressor p53’s specificity by bacterial redox protein azurin. Proc Natl Acad Sci USA 101: 4770–4775.    
  • 14. Punj V, Bhattacharyya S, Saint-dic D, et al. (2004) Bacterial cupredoxin azurin as an inducer of apoptosis and regression in human breast cancer. Oncogene 23: 2367–2378.    
  • 15. Bernardes N, Chakrabarty AM, Fialho AM (2013) Engineering of bacterial strains and their products for cancer therapy. Appl Microbiol Biotechnol 97: 5189–5199.    
  • 16. Apiyo D, Wittung-Stafshede P (2005) Unique complex between bacterial azurin and tumor-suppressor protein p53. Biochem Biophys Res Commun 332: 965–968.    
  • 17. Goto M, Yamada T, Kimbara K, et al. (2002) Induction of apoptosis in macrophages by Pseudomonas aeruginosa azurin: tumour-suppressor protein p53 and reactive oxygen species, but not redox activity, as critical elements in cytotoxicity. Mol Microbiol 47: 549–559.
  • 18. Yamada T, Fialho AM, Punj V, et al. (2005) Internalization of bacterial redox protein azurin in mammalian cells: entry domain and specificity. Cell Microbiol 7: 1418–1431.    
  • 19. Hong CS, Yamada T, Hashimoto W, et al. (2006) Disrupting the entry barrier and attacking brain tumors: The role of the Neisseria H.8 epitope and the Laz protein. Cell Cycle 5: 1633–1641.
  • 20. Chaudhari A, Mahfouz M, Fialho AM, et al. (2007) Cupredoxin-cancer interrelationship: azurin binding with EphB2, interference in EphB2 tyrosine phosphorylation and inhibition of cancer growth. Biochemistry 46: 1799–1810.    
  • 21. Mehta RR, Yamada T, Taylor BN, et al. (2011) A cell penetrating peptide derived from azurin inhibits angiogenesis and tumor growth by inhibiting phosphorylation of VEGFR-2, FAK and Akt. Angiogenesis 14: 355–369.    
  • 22. Mehta RR, Hawthorne M, Peng X, et al. (2010) A 28-amino-acid peptide fragment of the cupredoxin azurin prevents carcinogen-induced mouse mammary lesions. Cancer Prev Res 3: 1351–1360.    
  • 23. Warso MA, Richards JM, Mehta D, et al. (2013) A first-in-class, first-in-human, phase I trial of p28, a non-HDM2-mediated peptide inhibitor of p53 ubiquitination in patients with advanced solid tumours. Br J Cancer 108: 1061–1070.    
  • 24. Lulla RR, Goldman S, Yamada T, et al. (2016) Phase 1 trial of p28 (NSC745104), a non-HDM2-mediated peptide inhibitor of p53 ubiquitination in pediatric patients with recurrent or progressive central nervous system tumors: a pediatric brain tumor consortium study. Neuro Oncol pii: now047.
  • 25. Taylor BN, Mehta RR, Yamada T, et al. (2009) Noncationic peptides obtained from azurin preferentially enter cancer cells. Cancer Res 69: 537–546.    
  • 26. Jia L, Gorman GS, Coward LU, et al. (2011) Preclinical pharmacokinetics, metabolism, and toxicity of azurin-p28 (NSC745104) a peptide inhibitor of p53 ubiquitination. Cancer Chemother Pharmacol 68: 513–524.    
  • 27. Fialho AM, Bernardes N, Chakrabarty AM (2012) Recent patents on live bacteria and their products as potential anticancer agents. Recent Pat Anticancer Drug Discov 7: 31–55.    
  • 28. Bernardes N, Abreu S, Carvalho FA, et al. (2016) Modulation of membrane properties of lung cancer cells by azurin enhances the sensitivity to EGFR-targeted therapy and decreased β1 integrin-mediated adhesion. Cell Cycle 15: 1415–1424.    
  • 29. Yamada T, Das Gupta TK, Beattie CW (2016) p28-mediated activation of p53 in G2-M phase of the cell cycle enhances the efficacy of DNA damaging and antimitotic chemotherapy. Cancer Res 76: 2354–2365.
  • 30. Zaborina O, Dhiman N, Ling Chen M, et al. (2000) Secreted products of a nonmucoid Pseudomonas aeruginosa strain induce two modes of macrophage killing: external-ATP-dependent, P2Z-receptor-mediated necrosis and ATP-independent, caspase-mediated apoptosis. Microbiology 146: 2521–2530.    
  • 31. Bernardes N., Ribeiro AS., Abreu S, et al. (2013) The bacterial protein azurin impairs invasion and FAK/Src signaling in P-cadherin-overexpressing breast cancer cell models. PLoS One 8: e69023.    
  • 32. Ribeiro AS, Albergaria A, Sousa B, et al. (2010) Extracellular cleavage and shedding of P-cadherin: a mechanism underlying the invasive behavior of breast cancer cells. Oncogene 29: 392–402.    
  • 33. Bernardes N, Ribeiro AS, Abreu S, et al. (2014) High-throughput molecular profiling of a P-cadherin overexpressing breast cancer model reveals new targets for the anti-cancer bacterial protein azurin. Int J Biochem Cell Biol 50: 1–9.    
  • 34. Mollinedo F, de la Iglesia-Vicente J, Gajate C, et al. (2010) Lipid raft-targeted therapy in multiple myeloma. Oncogene 29: 3748–3757.    
  • 35. Coppari E, Yamada T, Bizzarri AR, et al. (2014) A nanotechnological molecular-modeling, and immunological approach to study the interaction of the anti-tumorigenic peptide p28 with the p53 family of proteins. Intl J Nanomedicine 9: 1799–1813.
  • 36. Yamada T, Das Gupta TK, Beattie CW (2013) P28, an anionic cell-penetrating peptide, increases the activity of wild type and mutated p53 without altering its conformation. Mol. Pharm 10: 3375–3383.
  • 37. Cho I, Blaser MJ (2012) The human microbiome: at the interface of health and disease. Nat Rev Genet 13: 260–270.
  • 38. Shaikh F, Abhinand P, Ragunath P (2012) Identification & characterization of Lactobacillus salavarius bacteriocins and its relevance in cancer therapeutics. Bioinformation 8: 589–594.    
  • 39. Nguyen C, Nguyen VD (2016) Discovery of azurin-like anticancer bacteriocins from human gut microbiome through homology modeling and molecular docking against the tumor suppressor p53. Biomed Res Int 2016: 8490482.
  • 40. Guex N, Peitsch MC (1997) SWISS-MODEL and the Swiss-PdbViewer: An environment for comparative protein modeling. Electrophoresis 18: 2714–2723.
  • 41. Gibrat JF, Madej T, Bryant SH (1996) Surprising similarities in structure comparison. Curr Opin Struct Biol 6: 377–385.    
  • 42. Paz I, Kligun E, Bengad B, et al. (2016) BindUP: a web server for non-homology-based prediction of DNA and RNA binding proteins. Nucleic Acids Res pii: gkw454.
  • 43. Jo S, Kim T, Iyer VG, et al. (2008) CHARMM-GUI: A web-based graphical user interface for CHARMM. J Comput Chem 29: 1859–1865.    


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Copyright Info: 2016, Arsenio M. Fialho, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

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