Cyanobacterial bioactive compound EMTAHDCA recovers splenomegaly, affects protein profile of E. coli and spleen of lymphoma bearing mice

  • Niveshika
  • Shashank Kumar Maurya
  • Balkrishna Tiwari
  • Sindhunath Chakraborty
  • Ekta Verma
  • Rajnikant Mishra
  • Arun Kumar MishraEmail author
Original Article


The antibacterial and anticancerous properties of EMTAHDCA have already been reported in our previous study. However, mode of action of EMTAHDCA is still elusive. The present study was aimed to investigate the molecular targets in Escherichia coli and spleen of lymphoma-bearing mice in response to cyanocompound 9-ethyliminomethyl-12 (morpholin-4-ylmethoxy)-5, 8, 13, 16-tetraaza -hexacene-2, 3- dicarboxylic acid (EMTAHDCA) isolated from fresh water cyanobacterium Nostoc sp. MGL001. Differential expressions of proteins were observed in both E. coli and spleen of lymphoma-bearing mice after EMTAHDCA treatment. In continuation of our previous study, the present study revealed that the antibacterial agent, EMTAHDCA causes the drastic reduction in synthesis of proteins related to replication, transcription, translation and transportation in E. coli. Probably the direct or indirect interaction of this compound with these important metabolic processes led to the reduction in growth and cell death. Furthermore, the anticancerous property of the compound EMTAHDCA reflected as down regulation in proteins of cell cycle, cellular metabolism, signalling, transcription and transport together with up regulation of apoptosis, DNA damage and immunoprotection related proteins in spleen of lymphoma-bearing mice. In this study the EMTAHDCA induced modulations in expression of proteins of key metabolic pathways in E. coli and spleen cells of lymphoma bearing mice helped in understanding the mechanism underlying the antibacterial and anti-cancerous property.


EMTAHDCA Drug target E. coli Mice 2DE gel electrophoresis MALDI TOF MS/MS 



The Head, Department of Botany, Banaras Hindu University, Varanasi, India is gratefully acknowledged for providing laboratory facilities. Ekta Verma is thankful to the UGC, New Delhi, Balkrishna Tiwari and Sindhunath Chakraborty are thankful to the ICAR AMAAS, New Delhi and Shashank Kumar Maurya is thankful to CSIR, New Delhi for financial support in the form of SRF.


Funding was provided by University Grants Commission (Grant Nos. Rajiv Gandhi National Fellowship-JRF and UGC-JRF), Council of Scientific and Industrial Research (Grant No. SRF), National Bureau of Agriculturally Important Microorganisms (Grant No. SRF).

Compliance with ethical standards

Conflict of interest

All the authors declare that they don’t have any conflict of interest.

Ethical approval

All the experiments were approved by the Animal Ethical Committee of Institute of Science, Banaras Hindu University, Varanasi 221005, India. IAE No. 1802/GO/RE/S/15/CPSEA.


  1. 1.
    Antoniou A, Pharoah PDP, Narod S, Risch HA, Eyfjord JE, Hopper JL, Loman N, Olsson H et al (2003) Average risks of breast and ovarian cancer associated with BRCA1 or BRCA2 mutations detected in case series unselected for family history: a combined analysis of 22 studies. Am J Hum Genet 72:1117–1130CrossRefGoogle Scholar
  2. 2.
    Bandow JE, Brötz H, Ole LI, Labischinski H, Hecker M (2003) Proteomic approach to understanding antibiotic action proteomic approach to understanding antibiotic action. Antimicrob Agents Chemother 47:948–955CrossRefGoogle Scholar
  3. 3.
    Basu B, Apte S (2012) Gamma radiation-induced proteome of Deinococcus radiodurans primarily targets DNA repair and oxidative stress alleviation. Mol Cell Prot 11: M111.011734CrossRefGoogle Scholar
  4. 4.
    Beaumont PO, Moore MJ, Ahmad K, Payne MM, Lee C, Riddick DS (1998) Role of glutathione S-transferases in the resistance of human colon cancer cell lines to doxorubicin. Cancer Res 58:947–955Google Scholar
  5. 5.
    Bharti B, Mishra R (2008) Lymphoma affects enzymes and protein profile of non-lymphatic tissues in mice. Int J Integra Biol 3:175–181Google Scholar
  6. 6.
    Bharti B, Mishra R (2011) Isoforms of Pax5 and co-regulation of T- and B-cells associated genes influence phenotypic traits of ascetic cells causing Dalton’s lymphoma. Biochim Biophys Acta 1813:2071–2078CrossRefGoogle Scholar
  7. 7.
    Bharti B, Mishra R (2015) Spleen-specific isoforms of Pax5 and Ataxin-7 as potential proteomic markers of lymphoma-affected spleen. Mol Cell Biochem 402:181–191CrossRefGoogle Scholar
  8. 8.
    Bullerjahn GS, Post AF (2014) Physiology and molecular biology of aquatic cyanobacteria. Front Microbiol 5:1–2CrossRefGoogle Scholar
  9. 9.
    Cabiscol E, Tamarit J, Ros J (2000) Oxidative stress in bacteria and protein damage by reactive oxygen species. Int Microbiol 3:3–8Google Scholar
  10. 10.
    Castielli O, De la Cerda B, Navarro JA, Hervás M, De la Rosa MA (2009) Proteomic analyses of the response of cyanobacteria to different stress conditions. FEBS Lett 583:1753–1758CrossRefGoogle Scholar
  11. 11.
    Chen J, Hu WJ, Wang C, Liu TW, Chalifour A, Chen J, Shen ZJ, Liu X, Wang WH, Zheng HL (2014) Proteomic analysis reveals differences in tolerance to acid rain in two broad-leaf tree species, Liquidambar formosana and Schima superba. PLoS ONE 9:1–15Google Scholar
  12. 12.
    Cragg GM, Newman DJ (2013) Natural products: a continuing source of novel drug leads. Biochim Biophys Acta 1830:3670–3695CrossRefGoogle Scholar
  13. 13.
    Dittmann E, Gugger M, Sivonen K, Fewer DP (2015) Natural products biosynthetic diversity and comparative genomics of the cyanobacteria. Trends Microbiol 23:642–652CrossRefGoogle Scholar
  14. 14.
    Dutta S, Ray S, Nagarajan K (2013) Glutamic acid as anticancer agent: an overview. Saudi Pharm 21:337–343CrossRefGoogle Scholar
  15. 15.
    Figueiredo J, Odete AB, Fardilha M (2014) Protein phosphatase 1 and its complexes in carcinogenesis. Curr Cancer Drug Targets 14:2–29CrossRefGoogle Scholar
  16. 16.
    Fridovich I (1995) Superoxide radical and superoxide dismutases. Annu Rev Biochem 64:97–112CrossRefGoogle Scholar
  17. 17.
    Henderson MC, Shaw YY, Wang H, Han H, Laurence H, Flynn G, Dorr RT, Von HDD (2009) UA62784; a novel inhibitor of CENP-E kinesin-like protein. Mol Cancer Ther 8:36–44CrossRefGoogle Scholar
  18. 18.
    Herbert B, Righetti PG (2000) A turning point in proteome analysis: sample prefractionation via. multicompartment electrolyzers with isoelectric membranes. Electrophoresis 21:3639–3648CrossRefGoogle Scholar
  19. 19.
    Hermann T (2005) Drugs targeting the ribosome. Curr Opin Struct Biol 15:355–366CrossRefGoogle Scholar
  20. 20.
    Imlay JA (2013) The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium. Nat Rev Microbiol 11:443–454CrossRefGoogle Scholar
  21. 21.
    Inoue J, Gohda J, Akiyama T, Semba K (2007) NF- κ B activation in development and progression of cancer. Cancer Sci 98:268–274CrossRefGoogle Scholar
  22. 22.
    Jen J, Wang Y (2016) Zinc finger proteins in cancer progression. J Biomed Sci 23:1–9CrossRefGoogle Scholar
  23. 23.
    Jesionek-Kupnicka D, Bojo M, Prochorec-Sobieszek M, Szumera-Ciec´kiewicz A, Jabłonska J, Kalinka-Warzocha E, Kordek R, Młynarski W et al (2016) HLA-G and MHC class II protein expression in diffuse large B-cell lymphoma. Arch Immunol Ther Exp 64:225–240CrossRefGoogle Scholar
  24. 24.
    Kaur M, Kaur T, Kamboj SS, Singh J (2016) Roles of Galectin-7 in Cancer Asian Pac. J Cancer Prev 17:455–461Google Scholar
  25. 25.
    Koiri RK, Trigun SK, Mishra L, Pandey K, Dixit D, Dubey SK (2009) Regression of Dalton’s lymphoma in vivo via decline in lactate dehydrogenase and induction of apoptosis by a ruthenium(II)-complex containing 4-carboxy N-ethylbenzamide as ligand. Invest New Drugs 6:503–516CrossRefGoogle Scholar
  26. 26.
    Li H, Lin X, Wang S, Peng X (2007) Identification and antibody therapeutic targeting of chloramphenicol-resistant outer membrane proteins in Escherichia coli. J Prot Res 6:3628–3636CrossRefGoogle Scholar
  27. 27.
    Lin J, Huang S, Zhang Q (2002) Outer membrane proteins: key players for bacterial adaptation in host niches. Microbes Infect 4:325–331CrossRefGoogle Scholar
  28. 28.
    Liu SK, Smith CA, Arnold R, Kiefer F, Mcglade CJ, Mcglade CJ (2000) The adaptor protein gads (Grb2-related adaptor downstream of Shc) is implicated in coupling hemopoietic progenitor kinase-1 to the activated TCR. J Immunol 165:1417–1426CrossRefGoogle Scholar
  29. 29.
    Louis KS, Siegel AC (2011) Cell viability analysis using trypan blue: manual and automated methods. Methods Mol Biol 740:7–12CrossRefGoogle Scholar
  30. 30.
    Lowry OH, Roseborugh NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  31. 31.
    Lu J, Pei H, Kaeck M, Thompson HJ (1997) Gene expression changes associated with chemically induced rat mammary carcinogenesis. Mol Carcinog 20:204–215CrossRefGoogle Scholar
  32. 32.
    Ma C, Sim S, Shi W, Du L, Xing D, Zhang Y (2009) Energy production genes sucB and ubiF are involved in persister survival and tolerance to multiple antibiotics and stresses in Escherichia coli. FEMS Microbiol Lett 303:33–40CrossRefGoogle Scholar
  33. 33.
    Malekinejad H, Bazargani-Gilani B, Tukmechi A, Ebrahimi H (2012) A cytotoxicity and comparative antibacterial study on the effect of Zatariamultiflora Boiss, Trachyspermum copticum essential oils and Enrofloxacinon Aeromonas hydrophila Avicenna. J Phytomed 2:188–195Google Scholar
  34. 34.
    Mistrik M, Bartek J (2010) Cyclin D3-dependent kinases in lymphoma: redundancy and implications for therapy. Cell Cycle 9:440–449CrossRefGoogle Scholar
  35. 35.
    Mosesso P, Piane M, Palitti F, Pepe G, Penna S, Chessa L (2005) The novel human gene aprataxin is directly involved in DNA single strand break repair. Cell Mol Life Sci 62:485–491CrossRefGoogle Scholar
  36. 36.
    Niveshika VE, Maurya SK, Mishra R, Mishra AK (2017) The combined use of in silico. in vitro, and in vivo analyses to assess anti-cancerous potential of a bioactive compound from cyanobacterium Nostoc sp. MGL001. Front Pharmacol. Google Scholar
  37. 37.
    Niveshika VE, Mishra AK, Singh AK, Singh VK (2016) Structural elucidation and molecular docking of a novel antibiotic compound from cyanobacterium Nostoc sp. MGL001. Front Microbiol. Google Scholar
  38. 38.
    Northey JJ, Dong Z, Ngan E, Kaplan A, Hardy WR, Pawson T, Siegel PM (2013) Distinct phosphotyrosine-dependent functions of the ShcA adaptor protein are required for transforming growth factor (TGF)-induced breast cancer cell migration, invasion and metastasis. J Biol Chem 288:5210–5222CrossRefGoogle Scholar
  39. 39.
    Pan JA, Sun Y, Jiang YP, Bott AJ, Jaber N, Dou Z, Yang B, Chen JS et al (2017) TRIM21 ubiquitylates SQSTM1/p62 and suppresses protein sequestration to regulate redox homeostasis. Mol Cell 61:720–733CrossRefGoogle Scholar
  40. 40.
    Rosenberg SM, Shee C, Frisch RL, Hastings PJ (2012) Stress-induced mutation via DNA breaks in Escherichia coli: a molecular mechanism with implications for evolution and medicine. BioEssays 34:885–892. CrossRefGoogle Scholar
  41. 41.
    Salvador-Reyesa LA, Luesch H (2015) Biological targets and mechanisms of action of natural products from marine cyanobacteria. Nat Prod Rep 32:478–503CrossRefGoogle Scholar
  42. 42.
    Shevchenko A, Tower H, Havlis J, Olsen JV, Mann M (2006) In gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1:2856–2860CrossRefGoogle Scholar
  43. 43.
    Singh S, Kate BN, Banerjee UC (2005) Bioactive compounds from cyanobacteria and microalgae: an overview. Crit Rev Biotechnol 25:73–95CrossRefGoogle Scholar
  44. 44.
    Thibodeau J, Bourgeois-Daigneault MC, Lapointe R (2012) Targeting the MHC Class II antigen presentation pathway in cancer immunotherapy. Onco Immunol 1:908–916Google Scholar
  45. 45.
    Townsend DM, Tew KD (2003) The role of glutathione- S -transferase in anti-cancer drug resistance. Oncogene 22:7369–7375CrossRefGoogle Scholar
  46. 46.
    Tripathi MK, Kumar M, Deepali S, Asthana RK, Nigam S (2017) Proteomic analysis of sensitive and resistant isolates of Escherichia coli in understanding target(s) of a cyanobacterial biomolecule Hapalindole-T. J Aqua Res Dev 8:1–5Google Scholar
  47. 47.
    Ulloa F, Gonz A, Meffre D, Barrecheguren PJ (2015) Blockade of the SNARE protein syntaxin 1 inhibits glioblastoma tumor growth. PLoS ONE 10:1–10CrossRefGoogle Scholar
  48. 48.
    VanBogelen RA, Schiller EE, Thomas JD, Neidhardt FC (1999) Diagnosis of cellular states of microbial organisms using proteomics. Electrophoresis 20:2149–2159CrossRefGoogle Scholar
  49. 49.
    Warburg O, Wind F, Negelein E (1927) The metabolism of tumors in the body. J Gen Physiol 8:519–530CrossRefGoogle Scholar
  50. 50.
    Wise DR, Thompson CB (2010) Glutamine addiction: a new therapeutic target in cancer. Trends Biochem Sci 35:427–433CrossRefGoogle Scholar
  51. 51.
    Xia Y, Shen S, Verma IM (2014) NF-kB, an active player in human cancers. Cancer Immunol Res 2:823–830CrossRefGoogle Scholar
  52. 52.
    Xu C, Lin X, Ren H, Zhang Y, Wang S, Peng X (2006) Analysis of outer membrane proteome of Escherichia coli related to resistance to ampicillin and tetracycline. Proteomics 6:462–473CrossRefGoogle Scholar
  53. 53.
    Yamamoto-furusho JK, Barnich N, Xavier R, Hisamatsu T, Podolsky DK (2006) Centaurin β1 down-regulates nucleotide-binding oligomerization domains 1- and 2-dependent, NF-kB activation. J Biol Chem 281:36060–36070CrossRefGoogle Scholar
  54. 54.
    Zhu X, Ding M, Yu ML, Feng MX, Tan LJ, Zhao FK (2010) Identification of galectin-7 as a potential biomarker for oesophageal squamous cell carcinoma by proteomic analysis. BMC Cancer 10:1–12CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Laboratory of Microbial Genetics, Department of BotanyBanaras Hindu UniversityVaranasiIndia
  2. 2.Biochemistry and Molecular Biology Laboratory, Department of ZoologyBanaras Hindu UniversityVaranasiIndia

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