Amino Acids

, Volume 32, Issue 1, pp 145–156 | Cite as

Focus on phosphohistidine

  • P. V. Attwood
  • M. J. Piggott
  • X. L. Zu
  • P. G. Besant
Review Article


Phosphohistidine has been identified as an enzymic intermediate in numerous biochemical reactions and plays a functional role in many regulatory pathways. Unlike the phosphoester bond of its cousins (phosphoserine, phosphothreonine and phosphotyrosine), the phosphoramidate (P–N) bond of phosphohistidine has a high ΔG° of hydrolysis and is unstable under acidic conditions. This acid-lability has meant that the study of protein histidine phosphorylation and the associated protein kinases has been slower to progress than other protein phosphorylation studies.

Histidine phosphorylation is a crucial component of cell signalling in prokaryotes and lower eukaryotes. It is also now becoming widely reported in mammalian signalling pathways and implicated in certain human disease states. This review covers the chemistry of phosphohistidine in terms of its isomeric forms and chemical derivatives, how they can be synthesized, purified, identified and the relative stabilities of each of these forms. Furthermore, we highlight how this chemistry relates to the role of phosphohistidine in its various biological functions.

Keywords: Phosphohistidine – Histidine kinase – Phosphoramidate 


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  1. Besant, PG, Attwood, PV 1998Problems with phosphoamino acid analysis using alkaline hydrolysisAnal Biochem265187190PubMedCrossRefGoogle Scholar
  2. Besant, PG, Attwood, PV 2000Detection of a mammalian histone H4 kinase that has yeast histidine kinase-like enzymic activityInt J Biochem Cell Biol32243253PubMedCrossRefGoogle Scholar
  3. Besant, PG, Attwood, PV 2005Mammalian histidine kinasesBiochim Biophys Acta1754281290PubMedGoogle Scholar
  4. Besant, PG, Byrne, L, Thomas, G, Attwood, PV 1998A chromatographic method for the preparative separation of phosphohistidinesAnal Biochem258372375PubMedCrossRefGoogle Scholar
  5. Besant, PG, Lasker, MV, Bui, CD, Turck, CW 2000Phosphohistidine analysis using reversed-phase thin-layer chromatographyAnal Biochem282149153PubMedCrossRefGoogle Scholar
  6. Besant, PG, Tan, E, Attwood, PV 2003Mammalian protein histidine kinasesInt J Biochem Cell Biol35297309PubMedCrossRefGoogle Scholar
  7. Bieber, LL, Boyer, PD 196632p-labeling of mitochondrial protein and lipid fractions and their relation to oxidative phosphorylationJ Biol Chem24153755383PubMedGoogle Scholar
  8. Carpenter, FH 1960Free-energy change in hydrolytic reactions: nonionized-compound conventionJ Am Chem Soc8211111122CrossRefGoogle Scholar
  9. Corbridge, DEC 1985Phosphorus, an outline of its chemistry, biochemistry and technologyStudies in inorganic chemistry 63ElsevierAmsterdam371Google Scholar
  10. Crovello, CS, Furie, BC, Furie, B 1995Histidine phosphorylation of P-selectin upon stimulation of human platelets: a novel pathway for activation-dependent signal transductionCell82279286PubMedCrossRefGoogle Scholar
  11. Deluca, M, Ebner, KE, Hultquist, DE, Kriel, G, Peter, JB, Moyer, RW, Boyer, PD 1963The Isolation and Identification of phosphohistidine from mitochondrial proteinBiochem Z338512525PubMedGoogle Scholar
  12. Dhillon, NK, Sharma, S, Khuller, GK 2003Signaling through protein kinases and transcriptional regulators in Candida albicansCrit Rev Microbiol29259275PubMedGoogle Scholar
  13. DiSabato, G, Jencks, WP 1961Mechanism and catalysis reactions of acyl phosphates. I Nucleophilic reactionsJ Am Chem Soc8343934400CrossRefGoogle Scholar
  14. Duclos, B, Marcandier, S, Cozzone, AJ 1991Chemical properties and separation of phosphoaminoacids by thin-layer chromatography and/or electrophoresisMethods Enzymol2101021Google Scholar
  15. Edlund, B 1982Effects of chemical modification of lysine, tyrosine and tryptophan residues in pea seed nucleoside diphosphate kinase and inhibition of the enzyme with antibodiesUps J Med Sci87251258PubMedCrossRefGoogle Scholar
  16. El-Maghrabi, MR, Pilkis, SJ 1984Rat liver 6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase: A review of relationships between the two activities of the enzymeJ Cell Biochem26117PubMedCrossRefGoogle Scholar
  17. Emsley, J, Hall, D 1976The chemistry of phosphorous; environmental, organic, inorganic, biochemical and spectroscopic aspectsHarper & RowLondon386Google Scholar
  18. Engel, M, Veron, M, Theisinger, B, Lacombe, ML, Seib, T, Dooley, S, Welter, C 1995A novel serine/threonine-specific protein phosphotransferase activity of nm23/nucleoside-diphosphate kinaseEur J Biochem234200207PubMedCrossRefGoogle Scholar
  19. Fujitaki, JM, Fung, G, Oh, EY, Smith, RA 1981Characterization of chemical and enzymatic acid-labile phosphorylation of histone h4 using phosphorus-31 nuclear magnetic resonanceBiochemistry2036583664PubMedCrossRefGoogle Scholar
  20. Gassner, M, Stehlik, D, Schrecker, O, Hengstenberg, W, Maurer, W, Ruterjans, H 1977The phosphoenolpyruvate-dependent phosphotransferase system of staphylococcus aureus. 2. 1 h and 31p-nuclear-magnetic-resonance studies on the phosphocarrier protein hpr, phosphohistidines and phosphorylated hprEur J Biochemis75287296CrossRefGoogle Scholar
  21. Grefen, C, Harter, K 2004Plant two-component systems: principles, functions, complexity and cross talkPlanta219733742PubMedCrossRefGoogle Scholar
  22. Gross, J, Rajavel, M, Segura, E, Grubmeyer, C 1996Energy coupling in salmonella typhimurium nicotinic acid phosphoribosyltransferase: Identification of his-219 as site of phosphorylationBiochemistry3539173924PubMedCrossRefGoogle Scholar
  23. Hartsough, MT, Morrison, DK, Salerno, M, Palmieri, D, Ouatas, T, Mair, M, Patrick, J, Steeg, PS 2002Nm23-h1 metastasis suppressor phosphorylation of kinase suppressor of ras via a histidine protein kinase pathwayJ Biol Chem2773238932399PubMedCrossRefGoogle Scholar
  24. Hays, JB, Simoni, RD, Roseman, S 1973Sugar transport. V. A trimeric lactose-specific phosphocarrier protein of the Staphylococcus aureus phosphotransferase systemJ Biol Chem248941956PubMedGoogle Scholar
  25. Hobbs, E, Corbridge, DEC, Raistrick, B 1953The crystal structure of monosodium phosphoramidate, NaHPO3NH2 Acta Crystallogr6621626CrossRefGoogle Scholar
  26. Huang, JM, Wei, YF, Kim, YH, Osterberg, L, Matthews, HR 1991Purification of a protein histidine kinase from the yeast saccharomyces cerevisiae. The first member of this class of protein kinasesJ Biol Chem26690239031PubMedGoogle Scholar
  27. Huebner, VD, Matthews, HR 1985Phosphorylation of histidine in proteins by a nuclear extract of physarum polycephalum plasmodiaJ Biol Chem2601610616113PubMedGoogle Scholar
  28. Hultquist, DE 1968The preparation and characterization of phosphorylated derivatives of histidineBiochim Biophys Acta153329340PubMedCrossRefGoogle Scholar
  29. Hultquist, DE, Moyer, RW, Boyer, PD 1966The preparation and characterization of 1-phosphohistidine and 3-phosphohistidineBiochemistry5322331PubMedCrossRefGoogle Scholar
  30. Jencks, WP, Gilchrist, M 1965Reactions of nucleophilic reagents with phosphoramidateJ Am Chem Soc8731993209CrossRefGoogle Scholar
  31. Kirby, AJ 1977The organic chemistry of phosphate transferPhosphorus in the environment: its chemistry and biochemistryElsevierAmsterdam117134Google Scholar
  32. Klumpp, S, Krieglstein, J 2002Phosphorylation and dephosphorylation of histidine residues in proteinsEur J Biochem26910671071PubMedCrossRefGoogle Scholar
  33. Klumpp, S, Krieglstein, J 2005Reversible phosphorylation of histidine residues in vertebrate proteinsBiochim Biophys Acta1754291295PubMedGoogle Scholar
  34. Kowluru, A 2002Identification and characterization of a novel protein histidine kinase in the islet beta cell: evidence for its regulation by mastoparan, an activator of G-proteins and insulin secretionBiochem Pharmacol6320912100PubMedCrossRefGoogle Scholar
  35. Kumble, KD, Ahn, K, Kornberg, A 1996Phosphohistidyl active sites in polyphosphate kinase of escherichia coliProc Natl Acad Sci USA931439114395PubMedCrossRefGoogle Scholar
  36. Lasker, MV, Bui, CD, Besant, PG, Sugawara, K, Thai, P, Medzihradszky, G, Turck, CW 1999Protein histidine phosphorylation: Increased stability of thiophosphohistidineProtein Sci821772185PubMedCrossRefGoogle Scholar
  37. Levene, PA, Alsberg, CL 1906The cleavage products of vitellinJ Biol Chem2127133Google Scholar
  38. Lipmann, FA, Levene, PA 1932Serinephosphoric acid obtained on hydrolysis of vitellinic acidJ Biol Chem98109114Google Scholar
  39. Lloyd, GJ, Hsu, C-M, Cooperman, BS 1970On the reactivity of phosphorylimidazole, an analog of known phosphorylated enzymesJ Am Chem Soc9348894892Google Scholar
  40. Lloyd, GJ, Hsu, C-M, Cooperman, BS 1971On the reactivity of phosphorylimidazole, an analog of known phosphorylated enzymesJ Am Chem Soc9348894892PubMedCrossRefGoogle Scholar
  41. Mahler, HR, Cordes, EH 1971Biological chemistry2Harper & RowNew York24Google Scholar
  42. Matthews, HR 1995Protein kinases and phosphatases that act on histidine, lysine, or arginine residues in eukaryotic proteins – a possible regulator of the mitogen-activated protein kinase cascadePharmacol Ther67323350PubMedCrossRefGoogle Scholar
  43. McLaughlin, AC, Leigh, JS,Jr, Cohn, M 1976Magnetic resonance study of the three-dimensional structure of creatine kinase-substrate complexesJ Biol Chem25127772787PubMedGoogle Scholar
  44. Modro TA (1981) Phosphoric and carboxylic amides; comparison of bonding and reactivity. ACS Symposium Series, 171 (Phosphorus Chemistry), 619–622Google Scholar
  45. Müller, Th, Rathlev, T, Rosenberg, Th 1956Special cases of nonenzymic transphosphorylationBiochim Biophys Acta19563564PubMedCrossRefGoogle Scholar
  46. Munoz-Dorado, J, Almaula, N, Inouye, S, Inouye, M 1993Autophosphorylation of nucleoside diphosphate kinase from myxococcus xanthusJ Bacteriol17511761181PubMedGoogle Scholar
  47. Napper, S, Kindrachuk, J, Olson, DJ, Ambrose, SJ, Dereniwsky, C, Ross, AR 2003Selective extraction and characterization of a histidine-phosphorylated peptide using immobilized copper(ii) ion affinity chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometryAnal Chem7517411747PubMedCrossRefGoogle Scholar
  48. Narindrasorasak, S, Bridger, WA 1977Phosphoenolypyruvate synthetase of escherichia coli: Molecular weight, subunit composition, and identification of phosphohistidine in phosphoenzyme intermediateJ Biol Chem25231213127PubMedGoogle Scholar
  49. Pirrung, MC 1999Histidine kinases and two-component signal transduction systemsChem Biol6167175CrossRefGoogle Scholar
  50. Pirrung, MC, James, KD, Rana, VS 2000Thiophosphorylation of histidineJ Org Chem6584488453PubMedCrossRefGoogle Scholar
  51. Ramirez, F, Okazaki, H, Marececk, JF 1977A cyclic enediol N-phosphorylpyrrole, and its contrasting behaviour vs the analogous N-PhosphorylimidazoleTetrahedron Lett1829272930CrossRefGoogle Scholar
  52. Schenkels, C, Erni, B, Reymond, JL 1999Phosphofurylalanine, a stable analog of phosphohistidineBioorg Med Chem Lett914431446PubMedCrossRefGoogle Scholar
  53. Severin, SE, Yudelovich, Rya 1947Synthesis and properties of phosphorylated β-alanine, l-histidine, and α-alanineBiokhimiya12105110Google Scholar
  54. Spronk, AM, Yoshida, H, Wood, HG 1976Isolation of 3-phosphohistidine from phosphorylated pyruvate, phosphate dikinaseProc Natl Acad Sci USA7344154419PubMedCrossRefGoogle Scholar
  55. Steeg, PS, Palmieri, D, Ouatas, T, Salerno, M 2003Histidine kinases and histidine phosphorylated proteins in mammalian cell biology, signal transduction and cancerCancer Lett190112PubMedCrossRefGoogle Scholar
  56. Stock, JB, Stock, AM, Mottonen, JM 1990Signal transduction in bacteriaNature344395400PubMedCrossRefGoogle Scholar
  57. Tan, E, Besant, PG, Attwood, PV 2002Mammalian histidine kinases: Do they really exist?Biochemistry4138433851PubMedCrossRefGoogle Scholar
  58. Tan, E, Besant, PG, Zu, XL, Turck, CW, Bogoyevitch, MA, Lim, SG, Attwood, PV, Yeoh, GC 2004Histone H4 histidine kinase displays the expression pattern of a liver oncodevelopmental markerCarcinogenesis2520832088PubMedCrossRefGoogle Scholar
  59. Tauler, A, El-Maghrabi, MR, Pilkis, SJ 1987Functional homology of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, phosphoglycerate mutase, and 2,3-bisphosphoglycerate mutaseJ Biol Chem2621680816815PubMedGoogle Scholar
  60. Wagner, PD, Steeg, PS, Vu, ND 1997Two-component kinase-like activity of nm23 correlates with its motility-suppressing activityProc Natl Acad Sci USA9490009005PubMedCrossRefGoogle Scholar
  61. Walinder, O 1969Evidence of the presence of 1-phosphohistidine as the main phosphohistidine as the main phosphorylated component at the active site of bovine liver nucleoside diphosphate kinaseActa Chem Scand23339341PubMedCrossRefGoogle Scholar
  62. Waygood, EB, Mattoo, RL, Peri, KG 1984Phosphoproteins and the phosphoenolpyruvate: sugar phosphotransferase system in Salmonella typhimurium and Escherichia coli: evidence for IIImannose, IIIfructose, IIIglucitol, and the phosphorylation of enzyme IImannitol and enzyme IIN-acetylglucosamineJ Cell Biochem25139159PubMedCrossRefGoogle Scholar
  63. Waygood, EB, Pasloske, K, Delbaere, LT, Deutscher, J, Hengstenberg, W 1988Characterization of the 1-phosphohistidinyl residue in the phosphocarrier protein hpr of the phosphoenolpyruvate: Sugar phosphotransferase system of streptococcus faecalisBiochem Cell Biol667680PubMedCrossRefGoogle Scholar
  64. Wind, M, Wegener, A, Kellner, R, Lehmann, WD 2005Analysis of chea histidine phosphorylation and its influence on protein stability by high-resolution element and electrospray mass spectrometryAnal Chem7719571962PubMedCrossRefGoogle Scholar
  65. Zetterqvist, O, Engstrom, L 1966Isolation of [32p]phosphohistidine from different rat-liver cell fractions after incubation with [32p]adenosine triphosphateBr J Haematol12520530Google Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • P. V. Attwood
    • 1
  • M. J. Piggott
    • 1
  • X. L. Zu
    • 1
  • P. G. Besant
    • 1
  1. 1.School of Biomedical, Biomolecular and Chemical SciencesThe University of Western AustraliaPerthAustralia

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