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Electrochemical determination of pyrophosphate at nanomolar levels using a gold electrode covered with a cysteine nanofilm and based on competitive coordination of Cu(II) ion to cysteine and pyrophosphate

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Abstract

We are presenting an electrochemical method for the determination of pyrophosphate ions (PPi) that is based on the competitive coordination of Cu(II) ion to a nanofilm of cysteine (Cys) and dissolved PPi. Cys was immobilized on the surface of a gold electrode by self-assembly. The Cys-modified gold electrode was loaded with Cu(II) ion which is released from the surface on addition of a sample containing PPi. The sensor shows an unprecedented electrochemical response to PPi, and the reduction peak currents is linearly related to the logarithm of the concentration of PPi in the 100 nM to 10 mM range (with an R2 or 0.982). The limit of detection is ~10 nM which is lower than the detection limits hitherto reported for PPi. Adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and common anions give a much weaker response. The method demonstrated here is simple, effective, highly sensitive, hardly interfered, and does not require the addition of a reagent. The method was applied to the determination of PPi in (spiked) serum samples.

Schematic illustration of the pyrophosphate sensing process.

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References

  1. Huang MS, Sage AP, Lu J, Demer LL, Tintut Y (2008) Phosphate and pyrophosphate mediate PKA-induced vascular cell calcification. Biochem Biophys Res Commun 374:553

    Article  CAS  Google Scholar 

  2. Mansurova SE, Deryabina OA (1989) A simple colorimetric assay method for pyrophosphate in the presence of a 1000-fold excess of orthophosphate: application of the method to the study of pyrophosphate metabolism in mitochondria. Anal Biochem 176:390

    Article  CAS  Google Scholar 

  3. Hirose M, Abe-Hashimoto J, Ogura K, Tahara H, Ide T, Yoshimura TJ (1997) A rapid, useful and quantitative method to measure telomerase activity by hybridization protection assay connected with a telomeric repeat amplification protocol. Cancer Res Clin Oncol 123:337

    Article  CAS  Google Scholar 

  4. Tsui FWL (2012) Genetics and mechanisms of crystal deposition in calcium pyrophosphate deposition disease. Curr Rheumatol Rep 14:155

    Article  CAS  Google Scholar 

  5. Caswell A, Guilland-Cumming DF, Hearn PR, McGuire MKB, Russell RGG (1983) Pathogenesis of chondrocalcinosis and pseudogout. Metabolism of inorganic pyrophosphate and production of calcium pyrophosphate dihydrate crystals. Ann Rheum Dis 42:27

    Article  CAS  Google Scholar 

  6. March JG, Simonet B, Grases F (2001) Determination of pyrophosphate in renal calculi and urine by means of an enzymatic method. Clin Chim Acta 314:187

    Article  CAS  Google Scholar 

  7. Aldakov D, Anzenbacher P, Aldakov D, Anzenbacher P Jr (2004) Sensing of aqueous phosphates by polymers with dual modes of signal transduction. J Am Chem Soc 126:4752

    Article  CAS  Google Scholar 

  8. Nishiyabu R, Anzenbacher P Jr (2005) Sensing of antipyretic carboxylates by simple chromogenic calix[4] pyrroles. J Am Chem Soc 127:8270

    Article  CAS  Google Scholar 

  9. Climent E, Casasus R, Marcos MD, Sancenon F, Soto J (2009) Colorimetric sensing of pyrophosphate in aqueous media using bis-functionalised silica surfaces. Dalton Trans 24:4806

  10. Ronaghi M, Uhlen M, Nyre’n PM (1998) A sequencing method based on real-time. Science 281:363

    Article  CAS  Google Scholar 

  11. Gunnlaugsson T, Davis AP, O’Brien JE, Glynn M (2002) Fluorescent sensing of pyrophosphate and bis-carboxylates with charge neutral PET chemosensors. Org Lett 4:2449

    Article  CAS  Google Scholar 

  12. Singh NJ, Jun EJ, Chellappan K, Thangadurai D, Chandran RP, Hwang I-C, Yoon J, Kim KS (2007) Quinoxaline-imidazolium receptors for unique sensing of pyrophosphate and acetate by charge transfer. Org Lett 9:485

    Article  CAS  Google Scholar 

  13. Aldakov D, Anzenbacher P (2003) Dipyrrolyl quinoxalines with extended chromophores are efficient fluorimetric sensors for pyrophosphate. Chem Commun 27:1394

  14. Spangler C, Schaeferling M, Wolfbeis OS (2008) Fluorescent probes for micro determination of inorganic phosphates and biophosphates. Microchim Acta 161:1

    Article  CAS  Google Scholar 

  15. Schaeferling M, Wolfbeis OS (2007) Europium tetracycline as a luminescent probe for nucleoside phosphates, and its application to the determination of kinase activity. Chem Eur J 13:4342

    Article  CAS  Google Scholar 

  16. Lee DH, Kim SY, Hong JI (2004) A Fluorescent pyrophosphate sensor with High selectivity over ATP in water. Angew Chem Int Ed 43:4777

    Article  CAS  Google Scholar 

  17. Lee HN, Xu Z, Kim SK, Swamy KMK, Kim Y, Kim SJ, Yoon J (2007) Pyrophosphate-selective fluorescent chemosensor at physiological pH: formation of a unique excimer upon addition of pyrophosphate. J Am Chem Soc 129:3828

  18. Park C, Hong J (2010) A new fluorescent sensor for the detection of pyrophosphate based on a tetraphenylethylene moiety. Tetrahedron Lett 51:1960

    Article  CAS  Google Scholar 

  19. Bhowmik S, Ghosh BN, Marjomaki V, Rissanen KJ (2014) Nanomolar pyrophosphate detection in water and in a self-assembled hydrogel of a simple terpyridine-Zn2+ complex. J Am Chem Soc 136:5543

    Article  CAS  Google Scholar 

  20. Villamil-Ramos R, Yatsimirsky AK (2011) Selective fluorometric detection of pyrophosphate by interaction with alizarin red S–dimethyltin(IV) complex. Chem Commun 47:2694

    Article  CAS  Google Scholar 

  21. Zhao X, Huang C (2010) A molecular logic gate for the highly selective recognition of pyrophosphate with a hypocrellin A-Zn(II) complex. Analyst 135:2853

    Article  CAS  Google Scholar 

  22. Sokkalingam P, Kim DS, Hwang H, Sessler JL, Lee C-H (2012) A dicationic calix[4] pyrrole derivative and its use for the selective recognition and displacement-based sensing of pyrophosphate. Chem Sci 3:1819

    Article  CAS  Google Scholar 

  23. Rao AS, Singha S, Choi W, Ahn KH (2012) Studies on acedan-based mononuclear zinc complexes toward selective fluorescent probes for pyrophosphate. Org Biomol Chem 10:8410

    Article  CAS  Google Scholar 

  24. Zhu W, Huang X, Guo Z, Wu X, Yu H, Tian H (2012) A novel NIR fluorescent turn-on sensor for the detection of pyrophosphate anion in complete water system. Chem Commun 48:1784

    Article  CAS  Google Scholar 

  25. Xu Z, Singh NJ, Lim J, Pan J, Kim HN, Park S, Kim KS, Yoon J (2009) Unique sandwich stacking of pyrene-adenine-pyrene for selective and ratiometric fluorescent sensing of ATP at physiological pH. J Am Chem Soc 131:15528

    Article  CAS  Google Scholar 

  26. Marino N, Ikotun OF, Julve M, Lloret F, Cano J, Doyle RP (2011) Pyrophosphate-mediated magnetic interactions in Cu(II) coordination complexes. Inorg Chem 50:378

    Article  CAS  Google Scholar 

  27. Lenz GR, Martell AE (1964) Metal chelates of some sulfur-containing amino acids. Biochemistry 3:745

    Article  CAS  Google Scholar 

  28. English JB, Martell AE, Motekaitis RJ, Murase I (1997) Molecular interaction of pyrophosphate with 1,13-dioxa-4,7,10,16,20,24-hexaazacyclohexacosane (OBISDIPEN) and its mononuclear and dinuclear copper(II) complexes. Inorg Chim Acta 258:183

    Article  CAS  Google Scholar 

  29. Deng J, Yu P, Yang L, Mao L (2013) Competitive coordination of Cu2+ between cysteine and pyrophosphate ion: toward sensitive and selective sensing of pyrophosphate ion in synovial fluid of arthritis patients. Anal Chem 85:2516

    Article  CAS  Google Scholar 

  30. Li X, Yu P, Yang L, Wang F, Mao L (2012) An electrochemical method for investigation of conformational flexibility of active sites of trametes versicolor laccase based on sensitive determination of copper ion with cysteine-modified electrodes. Anal Chem 84:9416

    CAS  Google Scholar 

  31. Qian Q, Su L, Yu P, Cheng H, Lin Y, Jin X, Mao L (2012) Ionic liquid-assisted preparation of laccase-based biocathodes with improved biocompatibility. J Phys Chem B 116:5185

    Article  CAS  Google Scholar 

  32. Li X, Liu Y, Zhu A, Luo Y, Deng Z, Tian Y (2010) Real-time electrochemical monitoring of cellular H2O2 integrated with in situ selective cultivation of living cells based on dual functional protein microarrays at Au−TiO2 surfaces. Anal Chem 82:6512

    Article  CAS  Google Scholar 

  33. Lust G, Seegmiller JE (1976) Intracellular pyrophosphate levels were determined by coupled enzymatic and fluorimetric assay. Clin Chim Acta 66:241

    Article  CAS  Google Scholar 

  34. DengJ JQ, Wang Y, Yang L, Yu P, Mao L (2013) Real-time colorimetric assay of inorganic pyrophosphatase activity based on reversibly competitive coordination of Cu2+ between cysteine and pyrophosphate ion. Anal Chem 85:9409

    Article  Google Scholar 

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Acknowledgments

This work was financially supported by National Natural Science Foundation (21375088), Scientific Research Project of Beijing Educational Committee (KM201410028006), Youth Talent Project of the Beijing Municipal Commission of Education (CIT&TCD201504072), Scientific Research Base Development Program of the Beijing Municipal Commission of Education and the 2013 Program of Scientific Research Foundation for the Returned Overseas Chinese Scholars of Beijing Municipality.

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Authors and Affiliations

  1. Department of Chemistry, Capital Normal University, Beijing, 100048, China

    Yuqing Lin, Linbo Li, Keqing Wang, Yunfei Ji & Hong Zou

  2. College of Chemistry and Materials Science, Anhui Normal University, Wuhu, 241000, China

    Lianglu Hu

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  1. Yuqing Lin
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  3. Linbo Li
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Correspondence to Yuqing Lin.

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Lin, Y., Hu, L., Li, L. et al. Electrochemical determination of pyrophosphate at nanomolar levels using a gold electrode covered with a cysteine nanofilm and based on competitive coordination of Cu(II) ion to cysteine and pyrophosphate. Microchim Acta 182, 2069–2075 (2015). https://doi.org/10.1007/s00604-014-1414-3

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  • DOI: https://doi.org/10.1007/s00604-014-1414-3

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