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Phosphate-Modified Nucleotides for Monitoring Enzyme Activity

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Abstract

Nucleotides modified at the terminal phosphate position have been proven to be interesting entities to study the activity of a variety of different protein classes. In this chapter, we present various types of modifications that were attached as reporter molecules to the phosphate chain of nucleotides and briefly describe the chemical reactions that are frequently used to synthesize them. Furthermore, we discuss a variety of applications of these molecules. Kinase activity, for instance, was studied by transfer of a phosphate modified with a reporter group to the target proteins. This allows not only studying the activity of kinases, but also identifying their target proteins. Moreover, kinases can also be directly labeled with a reporter at a conserved lysine using acyl-phosphate probes. Another important application for phosphate-modified nucleotides is the study of RNA and DNA polymerases. In this context, single-molecule sequencing is made possible using detection in zero-mode waveguides, nanopores or by a Förster resonance energy transfer (FRET)-based mechanism between the polymerase and a fluorophore-labeled nucleotide. Additionally, fluorogenic nucleotides that utilize an intramolecular interaction between a fluorophore and the nucleobase or an intramolecular FRET effect have been successfully developed to study a variety of different enzymes. Finally, also some novel techniques applying electron paramagnetic resonance (EPR)-based detection of nucleotide cleavage or the detection of the cleavage of fluorophosphates are discussed. Taken together, nucleotides modified at the terminal phosphate position have been applied to study the activity of a large diversity of proteins and are valuable tools to enhance the knowledge of biological systems.

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References

  1. Lipmann F (1941) Metabolic generation and utilization of phosphate bond energy. Adv Enzymol Rel S Bi 1:99–162

    CAS  Google Scholar 

  2. Masai H et al (2010) Eukaryotic chromosome DNA replication: where, when, and how? Annu Rev Biochem 79:89–130

    Article  CAS  Google Scholar 

  3. Kalia D et al (2013) Nucleotide, c-di-GMP, c-di-AMP, cGMP, cAMP, (p)ppGpp signaling in bacteria and implications in pathogenesis. Chem Soc Rev 42:305–341

    Article  CAS  Google Scholar 

  4. McLennan AG (2000) Dinucleoside polyphosphates—friend or foe? Pharmacol Ther 87:73–89

    Article  CAS  Google Scholar 

  5. Robinson GA, Butcher RW, Sutherland EW (1968) Cyclic AMP. Annu Rev Biochem 37:149–174

    Article  Google Scholar 

  6. Corrigan RM, Grundling A (2013) Cyclic di-AMP: another second messenger enters the fray. Nat Rev Microbiol 11:513–524

    Article  CAS  Google Scholar 

  7. Sims PA, Greenleaf WJ, Duan H, Xie XS (2011) Fluorogenic DNA sequencing in PDMS microreactors. Nat Methods 8:575–580

    Article  CAS  Google Scholar 

  8. Hacker SM, Mortensen F, Scheffner M, Marx A (2014) Selective monitoring of the enzymatic activity of the tumor suppressor Fhit. Angew Chem Int Ed 53:10247–10250

    Article  CAS  Google Scholar 

  9. Patricelli MP et al (2007) Functional interrogation of the kinome using nucleotide acyl phosphates. Biochem US 46:350–358

    Article  CAS  Google Scholar 

  10. Suwal S, Pflum MKH (2010) Phosphorylation-dependent kinase-substrate cross-linking. Angew Chem Int Ed 49:1627–1630

    Article  CAS  Google Scholar 

  11. Hiratsuka T (1983) New ribose-modified fluorescent analogs of adenine and guanine nucleotides available as substrates for various enzymes. Biochim Biophys Acta 742:496–508

    Article  CAS  Google Scholar 

  12. Green KD, Pflum MKH (2007) Kinase-catalyzed biotinylation for phosphoprotein detection. J Am Chem Soc 129:10–11

    Article  CAS  Google Scholar 

  13. Korlach J et al (2008) Long, processive enzymatic DNA synthesis using 100% dye-labeled terminal phosphate-linked nucleotides. Nucleosides Nucleotides Nucleic Acids 27:1072–1083

    Article  CAS  Google Scholar 

  14. Dhar G, Bhaduri A (1999) Synthesis and characterization of stacked and quenched uridine nucleotide fluorophores. J Biol Chem 274:14568–14572

    Article  CAS  Google Scholar 

  15. Hofer A et al (2015) A modular synthesis of modified phosphoanhydrides. Chem Eur J 21:10116–10122

    Article  CAS  Google Scholar 

  16. Allen JJ et al (2007) A semisynthetic epitope for kinase substrates. Nat Methods 4:511–516

    Article  CAS  Google Scholar 

  17. Baranowski MR et al (2015) Synthesis of fluorophosphate nucleotide analogues and their characterization as tools for 19F NMR studies. J Org Chem 80:3982–3997

    Article  CAS  Google Scholar 

  18. Song H, Kerman K, Kraatz H-B (2008) Electrochemical detection of kinase-catalyzed phosphorylation using ferrocene-conjugated ATP. Chem Commun 4:502–504

    Article  Google Scholar 

  19. Martić S, Labib M, Freeman D, Kraatz H-B (2011) Probing the role of the linker in ferrocene–ATP conjugates: monitoring protein kinase catalyzed phosphorylations electrochemically. Chem Eur J 17:6744–6752

    Article  CAS  Google Scholar 

  20. Lee SE et al (2009) Synthesis and reactivity of novel γ-phosphate modified ATP analogues. Bioorg Med Chem Lett 19:3804–3807

    Article  CAS  Google Scholar 

  21. Serdjukow S et al (2014) Synthesis of γ-labeled nucleoside 5′-triphosphates using click chemistry. Chem Commun 50:1861–1863

    Article  CAS  Google Scholar 

  22. Hacker SM, Mex M, Marx A (2012) Synthesis and stability of phosphate modified atp analogues. J Org Chem 77:10450–10454

    Article  CAS  Google Scholar 

  23. Ermert S et al (2016) Different enzymatic processing of gamma-phosphoramidate and gamma-phosphoester-modified ATP analogues. ChemBioChem. doi:10.1002/cbic.201600590

    Google Scholar 

  24. Draganescu A, Hodawadekar SC, Gee KR, Brenner C (2000) Fhit-nucleotide specificity probed with novel fluorescent and fluorogenic substrates. J Biol Chem 275:4555–4560

    Article  CAS  Google Scholar 

  25. Wanat P et al (2015) Ethynyl, 2-propynyl, and 3-butynyl C-phosphonate analogues of nucleoside di- and triphosphates: synthesis and reactivity in CuAAC. Org Lett 17:3062–3065

    Article  CAS  Google Scholar 

  26. Kumar S et al (2005) Terminal phosphate labeled nucleotides: synthesis, applications, and linker effect on incorporation by DNA polymerases. Nucleosides Nucleotides Nucleic Acids 24:401–408

    Article  CAS  Google Scholar 

  27. Sood A et al (2005) Terminal phosphate-labeled nucleotides with improved substrate properties for homogeneous nucleic acid assays. J Am Chem Soc 127:2394–2395

    Article  CAS  Google Scholar 

  28. Hampton A, Kappler F, Picker D (1982) Species- or isozyme-specific enzyme inhibitors. 4. Design of a two-site inhibitor of adenylate kinase with isozyme selectivity. J Med Chem 25:638–644

    Article  CAS  Google Scholar 

  29. Johnson TB, Coward JK (1987) Synthesis of oligophosphopeptides and related ATP γ-peptide esters as probes for cAMP-dependent protein kinase. J Org Chem 52:1771–1779

    Article  CAS  Google Scholar 

  30. Ratnakar SJ, Alexander V (2005) Synthesis and relaxivity studies of a gadolinium(III) complex of ATP-conjugated DO3A as a contrast enhancing agent for MRI. Eur J Inorg Chem 2005:3918–3927

    Article  CAS  Google Scholar 

  31. Hacker SM et al (2013) Fluorogenic ATP analogues for online monitoring of ATP consumption: observing ubiquitin activation in real time. Angew Chem Int Ed 52:11916–11919

    Article  CAS  Google Scholar 

  32. Kumar S et al (2012) PEG-labeled nucleotides and nanopore detection for single molecule DNA sequencing by synthesis. Sci Rep 2:684

    Google Scholar 

  33. Eid J et al (2009) Real-time DNA sequencing from single polymerase molecules. Science 323:133–138

    Article  CAS  Google Scholar 

  34. Kwon SW et al (2003) Selective enrichment of thiophosphorylated polypeptides as a tool for the analysis of protein phosphorylation. Mol Cell Proteom 2:242–247

    CAS  Google Scholar 

  35. Blethrow JD, Glavy JS, Morgan DO, Shokat KM (2008) Covalent capture of kinase-specific phosphopeptides reveals Cdk1-cyclin B substrates. Proc Natl Acad Sci USA 105:1442–1447

    Article  CAS  Google Scholar 

  36. Allen JJ, Lazerwith SE, Shokat KM (2005) Bio-orthogonal affinity purification of direct kinase substrates. J Am Chem Soc 127:5288–5289

    Article  CAS  Google Scholar 

  37. Bishop AC et al (2000) A chemical switch for inhibitor-sensitive alleles of any protein kinase. Nature 407:395–401

    Article  CAS  Google Scholar 

  38. Liu Y et al (1998) Engineering Src family protein kinases with unnatural nucleotide specificity. Chem Biol 5:91–101

    Article  CAS  Google Scholar 

  39. Fouda AE, Pflum MKH (2015) A cell-permeable ATP analogue for kinase-catalyzed biotinylation. Angew Chem Int Ed 54:9618–9621

    Article  CAS  Google Scholar 

  40. Senevirathne C et al (2016) The generality of kinase-catalyzed biotinylation. Bioorg Med Chem 24:12–19

    Article  CAS  Google Scholar 

  41. Senevirathne C, Pflum MKH (2013) Biotinylated phosphoproteins from kinase-catalyzed biotinylation are stable to phosphatases: implications for phosphoproteomics. ChemBioChem 14:381–387

    Article  CAS  Google Scholar 

  42. Kerman K, Chikae M, Yamamura S, Tamiya E (2007) Gold nanoparticle-based electrochemical detection of protein phosphorylation. Anal Chim Acta 588:26–33

    Article  CAS  Google Scholar 

  43. Wang Z, Lévy R, Fernig DG, Brust M (2006) Kinase-catalyzed modification of gold nanoparticles: a new approach to colorimetric kinase activity screening. J Am Chem Soc 128:2214–2215

    Article  CAS  Google Scholar 

  44. Wang Z, Lee J, Cossins AR, Brust M (2005) Microarray-based detection of protein binding and functionality by gold nanoparticle probes. Anal Chem 77:5770–5774

    Article  CAS  Google Scholar 

  45. Ma C, Yeung ES (2010) Highly sensitive detection of DNA phosphorylation by counting single nanoparticles. Anal Bioanal Chem 397:2279–2284

    Article  CAS  Google Scholar 

  46. Kerman K et al (2008) Peptide biosensors for the electrochemical measurement of protein kinase activity. Anal Chem 80:9395–9401

    Article  CAS  Google Scholar 

  47. Martić S, Rains MK, Freeman D, Kraatz H-B (2011) Use of 5′-γ-ferrocenyl adenosine triphosphate (Fc-ATP) bioconjugates having poly(ethylene glycol) spacers in kinase-catalyzed phosphorylations. Bioconjugate Chem 22:1663–1672

    Article  CAS  Google Scholar 

  48. Martić S et al (2012) Versatile strategy for biochemical, electrochemical and immunoarray detection of protein phosphorylations. J Am Chem Soc 134:17036–17045

    Article  CAS  Google Scholar 

  49. Wang N et al (2015) Clickable 5′-γ-ferrocenyl adenosine triphosphate bioconjugates in kinase-catalyzed phosphorylations. Chem Eur J 21:4988–4999

    Article  CAS  Google Scholar 

  50. Parang K, Kohn JA, Saldanha SA, Cole PA (2002) Development of photo-crosslinking reagents for protein kinase–substrate interactions. FEBS Lett 520:156–160

    Article  CAS  Google Scholar 

  51. Petrousseva IO et al (2003) A new approach to the synthesis of the 5′-end substituted oligonucleotides using T4 polynucleotide kinase and γ-amides of ATP bearing photoreactive groups. Dokl Biochem Biophys 389:114–117

    Article  CAS  Google Scholar 

  52. Green KD, Pflum MKH (2009) Exploring kinase cosubstrate promiscuity: monitoring kinase activity through dansylation. ChemBioChem 10:234–237

    Article  CAS  Google Scholar 

  53. Wilke KE, Francis S, Carlson EE (2012) Activity-based probe for histidine kinase signaling. J Am Chem Soc 134:9150–9153

    Article  CAS  Google Scholar 

  54. Freeman R, Finder T, Gill R, Willner I (2010) Probing protein kinase (CK2) and alkaline phosphatase with CdSe/ZnS quantum dots. Nano Lett 10:2192–2196

    Article  CAS  Google Scholar 

  55. Wang L-J, Yang Y, Zhang C-Y (2015) Phosphorylation-directed assembly of a single quantum dot based nanosensor for protein kinase assay. Anal Chem 87:4696–4703

    Article  CAS  Google Scholar 

  56. Xiao Y, Guo L, Jiang X, Wang Y (2013) Proteome-wide discovery and characterizations of nucleotide-binding proteins with affinity-labeled chemical probes. Anal Chem 85:3198–3206

    Article  CAS  Google Scholar 

  57. Qiu H, Wang Y (2007) Probing adenosine nucleotide-binding proteins with an affinity-labeled nucleotide probe and mass spectrometry. Anal Chem 79:5547–5556

    Article  CAS  Google Scholar 

  58. Xiao Y, Ji D, Guo L, Wang Y (2014) Comprehensive characterization of SGTP-binding proteins by orthogonal quantitative SGTP-affinity profiling and SGTP/GTP competition assays. Anal Chem 86:4550–4558

    Article  CAS  Google Scholar 

  59. Villamor JG et al (2013) Profiling protein kinases and other atp binding proteins in arabidopsis using Acyl-ATP probes. Mol Cell Proteomics 12:2481–2496

    Article  CAS  Google Scholar 

  60. Wolfe LM et al (2013) A chemical proteomics approach to profiling the ATP-binding Proteome of Mycobacterium tuberculosis. Mol Cell Proteom 12:1644–1660

    Article  CAS  Google Scholar 

  61. Adachi J et al (2014) Proteome-wide discovery of unknown ATP-binding proteins and kinase inhibitor target proteins using an ATP probe. J Proteome Res 13:5461–5470

    Article  CAS  Google Scholar 

  62. Nordin BE et al (2015) ATP acyl phosphate reactivity reveals native conformations of hsp90 paralogs and inhibitor target engagement. Biochem US 54:3024–3036

    Article  CAS  Google Scholar 

  63. Xiao Y, Guo L, Wang Y (2013) Isotope-coded atp probe for quantitative affinity profiling of ATP-binding proteins. Anal Chem 85:7478–7486

    Article  CAS  Google Scholar 

  64. Patricelli MP et al (2011) In situ kinase profiling reveals functionally relevant properties of native kinases. Chem Biol 18:699–710

    Article  CAS  Google Scholar 

  65. Kwiatkowski N et al (2014) Targeting transcription regulation in cancer with a covalent CDK7 inhibitor. Nature 511:616–620

    Article  CAS  Google Scholar 

  66. Deng X et al (2011) Characterization of a selective inhibitor of the parkinson’s disease kinase LRRK2. Nat Chem Biol 7:203–205

    Article  CAS  Google Scholar 

  67. Xie T et al (2014) Pharmacological targeting of the pseudokinase Her3. Nat Chem Biol 10:1006–1012

    Article  CAS  Google Scholar 

  68. Chen Z et al (2015) Fluorogenic sequencing using halogen-fluorescein-labeled nucleotides. ChemBioChem 16:1153–1157

    Article  CAS  Google Scholar 

  69. Vassiliou W et al (2000) Exploiting polymerase promiscuity: a simple colorimetric RNA polymerase assay. Virology 274:429–437

    Article  CAS  Google Scholar 

  70. Kore AR, Yang B, Srinivasan B (2014) Efficient synthesis of terminal 4-methylumbelliferyl labeled 5-fluoro-2′-deoxyuridine-5′-O-tetraphosphate (Um-PPPP-FdU): a potential probe for homogenous fluorescent assay. Tetrahedron Lett 55:4822–4825

    Article  CAS  Google Scholar 

  71. Kozlov M et al (2005) Homogeneous fluorescent assay for RNA polymerase. Anal Biochem 342:206–213

    Article  CAS  Google Scholar 

  72. Korlach J et al. (2010) Chapter 20 - Real-Time DNA Sequencing from Single Polymerase Molecules. In: Nils GW (ed) Method Enzymol, vol 472. Academic Press, pp 431-455

  73. Levene MJ et al (2003) Zero-mode waveguides for single-molecule analysis at high concentrations. Science 299:682–686

    Article  CAS  Google Scholar 

  74. Travers KJ et al (2010) A flexible and efficient template format for circular consensus sequencing and SNP detection. Nucleic Acids Res 38:e159

    Article  CAS  Google Scholar 

  75. Coupland P et al (2012) Direct sequencing of small genomes on the pacific biosciences RS without library preparation. Biotechniques 53:365–372

    Article  CAS  Google Scholar 

  76. Yang Y et al (2015) Quantitative and multiplexed DNA methylation analysis using long-read single-molecule real-time bisulfite sequencing (SMRT-BS). BMC Genom 16:1–11

    Article  CAS  Google Scholar 

  77. Flusberg BA et al (2010) Direct detection of DNA methylation during single-molecule, real-time sequencing. Nat Methods 7:461–465

    Article  CAS  Google Scholar 

  78. Murray IA et al (2012) The methylomes of six bacteria. Nucleic Acids Res 40:11450–11462

    Article  CAS  Google Scholar 

  79. Clark TA et al (2012) Characterization of DNA methyltransferase specificities using single-molecule. Real-time DNA sequencing. Nucleic Acids Res 40:e29

    Article  CAS  Google Scholar 

  80. Song C-X et al (2012) Sensitive and specific single-molecule sequencing of 5-hydroxymethylcytosine. Nat Methods 9:75–77

    Article  CAS  Google Scholar 

  81. Au KF et al (2013) Characterization of the human ESC transcriptome by hybrid sequencing. P Natl Acad Sci USA 110:E4821–E4830

    Article  CAS  Google Scholar 

  82. Sharon D, Tilgner H, Grubert F, Snyder M (2013) A single-molecule long-read survey of the human transcriptome. Nat Biotechnol 31:1009–1014

    Article  CAS  Google Scholar 

  83. Vilfan ID et al (2013) Analysis of RNA base modification and structural rearrangement by single-molecule real-time detection of reverse transcription. J Nanobiotechnol 11:8

    Article  CAS  Google Scholar 

  84. Saletore Y et al (2012) The birth of the epitranscriptome: deciphering the function of RNA modifications. Genome Biol 13:175

    Article  CAS  Google Scholar 

  85. Pennisi E (2010) Semiconductors inspire new sequencing technologies. Science 327:1190

    Article  CAS  Google Scholar 

  86. Munroe DJ, Harris TJR (2010) Third-generation sequencing fireworks at marco island. Nat Biotechnol 28:426–428

    Article  CAS  Google Scholar 

  87. Fuller CW et al (2016) Real-time single-molecule electronic DNA sequencing by synthesis using polymer-tagged nucleotides on a nanopore array. Proc Natl Acad Sci USA 113:5233–5238

    Article  CAS  Google Scholar 

  88. Yarbrough LR (1978) Synthesis and properties of a new fluorescent analog of ATP: adenosine-5′-triphosphoro-γ-1-(5-sulfonic acid) napthylamidate. Biochem Biophys Res Commun 81:35–41

    Article  CAS  Google Scholar 

  89. Kasprzyk R et al (2016) Acetylpyrene-labelled 7-methylguanine nucleotides: unusual fluorescence properties and application to decapping scavenger activity monitoring. Org Biomol Chem 14:3863–3868

    Article  CAS  Google Scholar 

  90. Schlageck JG, Baughman M, Yarbrough LR (1979) Spectroscopic techniques for study of phosphodiester bond formation by Escherichia coli RNA polymerase. J Biol Chem 254:12074–12077

    CAS  Google Scholar 

  91. Bhat J et al (2006) High-throughput screening of RNA polymerase inhibitors using a fluorescent UTP analog. J Biomol Screen 11:968–976

    Article  CAS  Google Scholar 

  92. Vadakkadathmeethal K et al (2007) Fluorescence-based adenylyl cyclase assay adaptable to high throughput screening. Comb Chem High Throughput Screen 10:289–298

    Article  CAS  Google Scholar 

  93. Jameson EE et al (2005) Real-time detection of basal and stimulated G protein GTPase activity using fluorescent GTP analogues. J Biol Chem 280:7712–7719

    Article  CAS  Google Scholar 

  94. Asensio AC, Oaknin S, Rotllán P (1999) Fluorimetric detection of enzymatic activity associated with the human tumor suppressor Fhit protein. BBA Protein Struct M 1432:396–400

    Article  CAS  Google Scholar 

  95. Asensio AC et al (2007) Biochemical analysis of ecto-nucleotide pyrophosphatase phosphodiesterase activity in brain membranes indicates involvement of NPP1 isoenzyme in extracellular hydrolysis of diadenosine polyphosphates in central nervous system. Neurochem Int 50:581–590

    Article  CAS  Google Scholar 

  96. Oaknin S et al (2008) Binding of 5′-O-(2-Thiodiphosphate) to rat brain membranes is prevented by diadenosine tetraphosphate and correlates with ecto-nucleotide pyrophosphatase phosphodiesterase 1 (NPP1) activity. Neurosci Lett 432:25–29

    Article  CAS  Google Scholar 

  97. Wierzchowski J, Sierakowska H, Shugar D (1985) Continuous fluorimetric assay of nucleotide pyrophosphatase. kinetics, inhibitors, and extension to dinucleoside oligophosphatases. BBA Protein Struct Mol Enzymol 828:109–115

    Article  CAS  Google Scholar 

  98. Tolman GL, Barrio JR, Leonard NJ (1974) Chloroacetaldehyde-modified dinucleoside phosphates. Dynamic fluorescence quenching and quenching due to intramolecular complexation. Biochem US 13:4869–4878

    Article  CAS  Google Scholar 

  99. Ramos A, Rotllán P (1995) Specific dinucleoside polyphosphate cleaving enzymes from chromaffin cells: a fluorimetric study. BBA Protein Struct Mol Enzymol 1253:103–111

    Article  Google Scholar 

  100. Rotilán P et al (1991) Di (1, N6-ethenoadenosine)5′, 5‴-P1, P4-tetraphosphate, a fluorescent enzymatically active derivative of Ap4A. FEBS Lett 280:371–374

    Article  Google Scholar 

  101. Hardt N, Hacker SM, Marx A (2013) Synthesis and fluorescence characteristics of ATP-based FRET probes. Org Biomol Chem 11:8298–8305

    Article  CAS  Google Scholar 

  102. Hacker SM et al (2013) Fingerprinting differential active site constraints of ATPases. Chem Sci 4:1588–1596

    Article  CAS  Google Scholar 

  103. Hacker SM, Buntz A, Zumbusch A, Marx A (2015) Direct monitoring of nucleotide turnover in human cell extracts and cells by fluorogenic ATP analogs. ACS Chem Biol 10:2544–2552

    Article  CAS  Google Scholar 

  104. Gutiérrez Acosta OB et al (2014) Thiamine pyrophosphate stimulates acetone activation by desulfococcus biacutus as monitored by a fluorogenic ATP analogue. ACS Chem Biol 9:1263–1266

    Article  CAS  Google Scholar 

  105. Hacker SM, Hintze C, Marx A, Drescher M (2014) Monitoring enzymatic ATP hydrolysis by EPR spectroscopy. Chem Commun 50:7262–7264

    Article  CAS  Google Scholar 

  106. Baranowski MR, Nowicka A, Jemielity J, Kowalska J (2016) A fluorescent HTS assay for phosphohydrolases based on nucleoside 5′-fluorophosphates: its application in screening for inhibitors of mRNA decapping scavenger and PDE-I. Org Biomol Chem 14:4595–4604

    Article  CAS  Google Scholar 

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Acknowledgements

Financial support by the Deutsche Forschungsgemeinschaft (Grant SFB 969) and the Konstanz Research School Chemical Biology is gratefully acknowledged. S.M.H. also acknowledges the Deutsche Forschungsgemeinschaft, the Studienstiftung des deutschen Volkes and the Zukunftskolleg of the University of Konstanz for stipends.

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  1. Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstraße 10, 78464, Konstanz, Germany

    Susanne Ermert, Andreas Marx & Stephan M. Hacker

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  1. Susanne Ermert
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  2. Andreas Marx
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  3. Stephan M. Hacker
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Correspondence to Stephan M. Hacker.

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This article is part of the Topical Collection “Phosphate Labeling and Sensing in Chemical Biology”; edited by Henning Jessen.

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Ermert, S., Marx, A. & Hacker, S.M. Phosphate-Modified Nucleotides for Monitoring Enzyme Activity. Top Curr Chem (Z) 375, 28 (2017). https://doi.org/10.1007/s41061-017-0117-8

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