Advertisement

Synthesis of Phosphopeptides in the Fmoc Mode

  • Troy J. Attard
  • Neil O’Brien-Simpson
  • Eric C. ReynoldsEmail author
Special Issue: Peptides in Oral and Dental Research

Abstract

The synthesis of phosphopeptides has played a major role in the characterization of protein phosphorylation/dephosphorylation. The current range of synthesis protocols available provides a variety of possible routes by which to approach specific synthetic challenges, and this review article discusses these methods for the preparation of phosphopeptides and provides synthesis notes for each method. Phosphopeptide synthesis is achieved by either introduction of the phosphate group via post-synthetic (‘global’) phosphorylation of a resin-bound peptide or the incorporation of a pre-phosphorylated derivative into the growing peptide chain. Protocols and synthesis notes are provided for the synthesis of phosphoramidites, phosphotyrosyl, -seryl and -threonyl peptides and their mimetics, including thiophosphopeptides. The aim of this review was to provide a synthesis reference guide for Fmoc-based synthesis of both singly and multiply phosphorylated peptides, with particular emphasis given to the most successful and generally applicable methods.

Keywords

Phosphopeptide Phosphoramidite Global phosphorylation H-phosphonate Dialkyl phosphate Monobenzyl phosphate Difluoromethyl phosphonate Thiophosphopeptide 

Notes

Acknowledgements

This work was supported by the National Health and Medical Research Council grant 251707 and The CRC for Oral Health Science.

References

  1. Bannwarth W, Kitas EA (1992) Synthesis of multi-O4-phospho-L-tyrosine-containing peptides. Helv Chim Acta 75:707–714CrossRefGoogle Scholar
  2. Bannwarth W, Kueng E, Vorherr T (1996) Global phosphorylation of peptides containing oxidation-sensitive amino acids. Bioorg Med Chem Lett 6:2141–2146CrossRefGoogle Scholar
  3. Berkowitz DB, Eggen M, Shen Q, Shoemaker RK (1996) Ready access to fluorinated phosphonate mimics of secondary phosphates. Synthesis of the (a,a-difluoroalkyl)phosphonate analogs of L-phosphoserine, L-phosphoallothreonine, and L-phosphothreonine. J Org Chem 61:4666–4675PubMedCrossRefGoogle Scholar
  4. Burke TR Jr, Lee K (2003) Phosphotyrosyl Mimetics in the Development of Signal Transduction Inhibitors. Acc Chem Res 36:426–433PubMedCrossRefGoogle Scholar
  5. Burke TR Jr, Smyth MS, Nomizu M, Otaka A, Roller PR (1993) Preparation of fluoro- and hydroxy-4-(phosphonomethyl)-D,L-phenylalanine suitably protected for solid-phase synthesis of peptides containing hydrolytically stable analogs of O-phosphotyrosine. J Org Chem 58:1336–1340CrossRefGoogle Scholar
  6. Burke TR Jr, Kole HK, Roller PP (1994a) Potent inhibition of insulin receptor dephosphorylation by a hexamer peptide containing the phosphotyrosyl mimetic F2Pmp. Biochem Biophys Res Commun 204:129–134PubMedCrossRefGoogle Scholar
  7. Burke TR Jr, Smyth MS, Otaka A, Nomizu M, Roller PP, Wolf G, Case R, Shoelson SE (1994b) Nonhydrolyzable phosphotyrosyl mimetics for the preparation of phosphatase-resistant SH2 domain inhibitors. Biochemistry 33:6490–6494PubMedCrossRefGoogle Scholar
  8. Chao H-G, Bernatowicz MS, Reiss PD, Matsueda GR (1994) Synthesis and application of bis-silylethyl-derived phosphate-protected Fmoc-phosphotyrosine derivatives for peptide synthesis. J Org Chem 59:6687–6691CrossRefGoogle Scholar
  9. Chao H-G, Leiting B, Reiss PD, Burkhardt AL, Klimas CE, Bolen JB, Matsueda GR (1995) Synthesis and application of Fmoc-O-[Bis(dimethylamino)phosphono]tyrosine, a versatile protected phosphotyrosine equivalent. J Org Chem 60:7710–7711CrossRefGoogle Scholar
  10. Chen L, Wu L, Otaka A, Smyth MS, Roller PP, Burke TR Jr, Den Hertog J, Zhang Z-Y (1995) Why is phosphonodifluoromethyl phenylalanine a more potent inhibitory moiety than phosphonomethyl phenylalanine toward protein-tyrosine phosphatases? Biochem Biophys Res Commun 216:976–984PubMedCrossRefGoogle Scholar
  11. Cho H, Ramer SE, Itoh M, Winkler DG, Kitas E, Bannwarth W, Burn P, Saito H, Walsh CT (1991) Purification and characterization of a soluble catalytic fragment of the human transmembrane leukocyte antigen related (LAR) protein tyrosine phosphatase from an E. coli expression system. Biochemistry 30:6210–6216PubMedCrossRefGoogle Scholar
  12. Cohen P (2001) The role of protein phosphorylation in human health and disease. The Sir Hans Krebs Medal Lecture. Eur J Biochem 268:5001–5010PubMedCrossRefGoogle Scholar
  13. Coleman DR, Ren Z, Mandal PK, Cameron AG, Dyer GA, Muranjan S, Campbell M, Chen X, Mcmurray JS (2005) Investigation of the binding determinants of phosphopeptides targeted to the Src homology 2 domain of the signal transducer and activator of transcription 3. Development of a high-affinity peptide inhibitor. J Med Chem 48:6661–6670PubMedCrossRefGoogle Scholar
  14. Cross KJ, Huq NL, Palamara JE, Perich JW, Reynolds EC (2005a) Physicochemical characterization of casein phosphopeptide-amorphous calcium phosphate nanocomplexes. J Biol Chem 280:15362–15369PubMedCrossRefGoogle Scholar
  15. Cross KJ, Huq NL, Reynolds EC (2005b) Protein dynamics of bovine dentin phosphophoryn. J Pept Res 66:59–67PubMedCrossRefGoogle Scholar
  16. De Bont HBA, Van Boom JH, Liskamp RMJ (1990) Automatic synthesis of phosphopeptides by phosphorylation on the solid phase. Tetrahedron Lett 31:2497–2500CrossRefGoogle Scholar
  17. De Bont HBA, Van Boom JH, Liskamp RMJ (1991) Automatic synthesis of phosphopeptides on the solid phase. Pept. 1990, Proc Eur Pept Symp 21st, pp. 443–445Google Scholar
  18. De Bont DBA, Moree WJ, Van Boom JH, Liskamp RMJ (1993) Solid-phase synthesis of O-phosphorothioylserine- and -threonine-containing peptides as well as of O-phosphoserine- and -threonine-containing peptides. J Org Chem 58:1309–1317CrossRefGoogle Scholar
  19. Delange RJ, Kemp RG, Riley WD, Cooper RA, Krebs EG (1968) Activation of skeletal muscle phosphorylase kinase by adenosine triphosphate and adenosine 3′,5′-monophosphate. J Biol Chem 243:2200–2208PubMedGoogle Scholar
  20. Domchek SM, Auger KR, Chatterjee S, Burke TR Jr, Shoelson SE (1992) Inhibition of SH2 domain/phosphoprotein association by a nonhydrolyzable phosphonopeptide. Biochemistry 31:9865–9870PubMedCrossRefGoogle Scholar
  21. Fischer EH, Krebs EG (1955) Conversion of phosphorylase b to phosphorylase a in muscle extracts. J Biol Chem 216:121–132PubMedGoogle Scholar
  22. Flick MB, Sapi E, Perrotta PL, Maher MG, Halaban R, Carter D, Kacinski BM (1997) Recognition of activated CSF-1 receptor in breast carcinomas by a tyrosine 723 phosphospecific antibody. Oncogene 14:2553–2561PubMedCrossRefGoogle Scholar
  23. Fretz H (1997) Na-Fmoc-O,O-(dimethylphospho)-L-tyrosine fluoride: a convenient building block for the solid-phase synthesis of phosphotyrosyl peptides. Lett Pept Sci 4:171–176Google Scholar
  24. Fujii N, Otaka A, Sugiyama N, Hatano M, Yajima H (1987) Studies on peptides. CLV. Evaluation of trimethylsilyl bromide as a hard-acid deprotecting reagent in peptide synthesis. Chem Pharm Bull 35:3880–3883PubMedGoogle Scholar
  25. Garcia-Echeverria C (1995) Potential pyrophosphate formation upon use of Na-Fmoc-Tyr(PO3H2)-OH in solid-phase peptide synthesis. Lett Pept Sci 2:93–98CrossRefGoogle Scholar
  26. Garcia-Echeverria C (1996) Evaluation of coupling conditions for the incorporation of Na-Fmoc-Tyr(PO3H2)-OH in solid-phase peptide synthesis. Lett Pept Sci 2:369–373CrossRefGoogle Scholar
  27. Gerster M, Bleicher K, Bayer E (1996) Comparison of several sulfurizing reagents for synthesis of phosphorothioate oligonucleotides on different supports. Innovation and Perspectives in solid phase synthesis & combinatorial libraries: peptides, proteins and nucleic acids–small molecule organic chemical diversity. Collected papers, international symposium, 4th, Edinburgh, 12–16 September 1995, pp. 377–380Google Scholar
  28. Gordeev MF, Patel DV, Barker PL, Gordon EM (1994) N-a-Fmoc-4-phosphono(difluoromethyl)-L-phenylalanine: a new O-phosphotyrosine isosteric building block suitable for direct incorporation into peptides. Tetrahedron Lett 35:7585–7588CrossRefGoogle Scholar
  29. Green OM (1994) A rapid dealkylation of phosphonate diester for the preparation of 4-phosphonomethylphenylalanine-containing peptides. Tetrahedron Lett 35:8081–8084CrossRefGoogle Scholar
  30. Hamilton R, Shute RE, Travers J, Walker B, Walker BJ (1994) A convenient synthesis of phosphonate isosteres of serine phosphates. Tetrahedron Lett 35:3597–3600CrossRefGoogle Scholar
  31. Handa BK, Hobbs CJ (1998) An efficient and convenient procedure for the synthesis of N(alpha)-Fmoc-O-monobenzyl phosphonotyrosine. J Pept Sci 4:138–141PubMedCrossRefGoogle Scholar
  32. Herbst JJ, Andrews GC, Contillo LG, Singleton DH, Genereux PE, Gibbs EM, Lienhard GE (1995) Effect of the activation of phosphatidylinositol 3-kinase by a thiophosphotyrosine peptide on glucose transport in 3T3-L1 adipocytes. J Biol Chem 270:26000–26005PubMedCrossRefGoogle Scholar
  33. Higashimoto Y, Saito SI, Tong X-H, Hong A, Sakaguchi K, Appella E, Anderson CW (2000) Human p53 is phosphorylated on serines 6 and 9 in response to DNA damage-inducing agents. J Biol Chem 275:23199–23203PubMedCrossRefGoogle Scholar
  34. Hoffmann R, Wachs WO, Berger RG, Kalbitzer HR, Waidelich D, Bayer E, Wagner-Redeker W, Zeppezauer M (1995) Chemical phosphorylation of the peptides GGXA (X = S, T, Y): an evaluation of different chemical approaches. Int J Pept Protein Res 45:26–34PubMedCrossRefGoogle Scholar
  35. Hoffmann R, Lee VM, Leight S, Varga I, Otvos L Jr (1997) Unique Alzheimer’s disease paired helical filament specific epitopes involve double phosphorylation at specific sites. Biochemistry 36:8114–8124PubMedCrossRefGoogle Scholar
  36. Huq NL, Cross KJ, Talbo GH, Riley PF, Loganathan A, Crossley MA, Perich JW, Reynolds EC (2000) N-terminal sequence analysis of bovine dentin phosphophoryn after conversion of phosphoseryl to S-propylcysteinyl residues. J Dent Res 79:1914–1919PubMedCrossRefGoogle Scholar
  37. Imhof D, Nothmann D, Zoda MS, Hampel K, Wegert J, Boehmer FD, Reissmann S (2005) Synthesis of linear and cyclic phosphopeptides as ligands for the N-terminal SH2-domain of protein tyrosine phosphatase SHP-1. J Pept Sci 11:390–400PubMedCrossRefGoogle Scholar
  38. Johnson T, Packman LC, Hyde CB, Owen D, Quibell M (1996) Backbone protection and its application to the synthesis of a difficult phosphopeptide sequence. J Chem Soc Perkin Trans 1:719–728CrossRefGoogle Scholar
  39. Johnson TM, Perich JW, Bjorge JD, Fujita DJ, Cheng H-C (1997) Common and differential recognition of structural features in synthetic peptides by the catalytic domain and the Src-homology 2 (SH2) domain of pp60c-src. J Pept Res 50:365–371PubMedCrossRefGoogle Scholar
  40. Kitas EA, Perich JW, Wade JD, Johns RB, Tregear GW (1989) Fmoc-polyamide solid phase synthesis of an o-phosphotyrosine-containing tridecapeptide. Tetrahedron Lett 30:6229–6232CrossRefGoogle Scholar
  41. Kitas EA, Knorr R, Trzeciak A, Bannwarth W (1991a) Alternative strategies for the Fmoc solid-phase synthesis of O4-phospho-L-tyrosine-containing peptides. Helv Chim Acta 74:1314–1328CrossRefGoogle Scholar
  42. Kitas EA, Wade JD, Johns RB, Perich JW, Tregear GW (1991b) Preparation and use of Na-fluorenylmethoxycarbonyl-O-dibenzylphosphono-L-tyrosine in continuous flow solid phase peptide synthesis. J Chem Soc Chem Commun 338–339Google Scholar
  43. Kitas E, Kueng E, Bannwarth W (1994) Chemical synthesis of O-thiophosphotyrosyl peptides. Int J Pept Protein Res 43:146–153PubMedGoogle Scholar
  44. Krebs EG, Love DS, Bratvold GE, Trayser KA, Meyer WL, Fischer EH (1964) Purification and properties of rabbit skeletal muscle phosphorylase B kinase. Biochemistry 3:1022–1033PubMedCrossRefGoogle Scholar
  45. Lacombe JM, Andriamanampisoa F, Pavia AA (1990) Solid-phase synthesis of peptides containing phosphoserine using phosphate tert.-butyl protecting group. Int J Pept Protein Res 36:275–280PubMedCrossRefGoogle Scholar
  46. Levene PA, Alsberg C (1901) Paranucleic acid. Z Physiol Chem 31:543Google Scholar
  47. Luo S-Z, Li Y-M, Chen Z-Z, Abe H, Cui L-P, Nakanishi H, Qin X-R, Zhao Y-F (2003) Synthesis and matrix assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry study of phosphopeptide. Lett Pept Sci 10:57–62Google Scholar
  48. Meutermans WDF, Alewood PF (1996) A simple and effective procedure for the synthesis of the ‘difficult’ phosphotyrosine-containing peptide Stat 91 (695–708). Tetrahedron Lett 37:4765–4766CrossRefGoogle Scholar
  49. Mostafavi H, Austermann S, Forssmann WG, Adermann K (1996) Synthesis of phospho-urodilatin by combination of global phosphorylation with the segment coupling approach. Int J Pept Protein Res 48:200–207PubMedCrossRefGoogle Scholar
  50. Otaka A, Burke TR Jr, Smyth MS, Nomizu M, Roller PP (1993) Deprotection and cleavage methods for protected peptide resins containing 4-[(diethylphosphono)difluoromethyl]-DL-phenylalanine residues. Tetrahedron Lett 34:7039–7042CrossRefGoogle Scholar
  51. Ottinger EA, Shekels LL, Bernlohr DA, Barany G (1993) Synthesis of phosphotyrosine-containing peptides and their use as substrates for protein tyrosine phosphatases. Biochemistry 32:4354–4361PubMedCrossRefGoogle Scholar
  52. Otvos L Jr, Elekes I, Lee VM (1989) Solid-phase synthesis of phosphopeptides. Int J Pept Protein Res 34:129–133PubMedCrossRefGoogle Scholar
  53. Perich JW (1998) Synthesis of phosphopeptides via global phosphorylation on the solid phase: resolution of H-phosphonate formation. Lett Pept Sci 5:49–55Google Scholar
  54. Perich, JW, Johns RB (1988) Di-tert-butyl N,N-diethylphosphoramidite. A new phosphitylating agent for the efficient phosphorylation of alcohols. Synthesis 142–144Google Scholar
  55. Perich, JW, Johns RB (1991) Synthesis of casein-related peptides and phosphopeptides. X. A modified method for the synthesis of Ser(P)-containing peptides through 4-bromobenzyl phosphate protection. Aust J Chem 44:503Google Scholar
  56. Perich JW, Reynolds EC (1991a) The facile one-pot synthesis of Na-(9-fluorenylmethoxycarbonyl)-O-(O′,O″-dialkylphosphoro)-L-tyrosines using dialkyl N,N-diethylphosphoramidites. Synlett 577–578Google Scholar
  57. Perich JW, Reynolds EC (1991b) Fmoc/solid-phase synthesis of Tyr(P)-containing peptides through t-butyl phosphate protection. Int J Pept Protein Res 37:572–575PubMedCrossRefGoogle Scholar
  58. Perich JW, Nguyen Dung L, Reynolds EC (1991) The facile synthesis of Ala-Glu-Tyr(P)-Ser-Ala by global di-tert-butyl N,N-diethylphosphoramidite phosphite-triester phosphorylation of a resin-bound peptide. Tetrahedron Lett 32:4033–4034CrossRefGoogle Scholar
  59. Perich JW, Liepa I, Chaffee AL, Johns RB (1994) Fast atom bombardment mass spectra of some Na-(t-butoxycarbonyl)-O-(diorganylphosphono)-L-serines and O-(diorganylphosphono)seryl-containing dipeptides and tripeptides. Aust J Chem 47:229–245CrossRefGoogle Scholar
  60. Perich JW, Liepa I, Chaffee AL, Johns RB (1996a) The analysis of multiple O-phosphoseryl-containing peptides by fast-atom-bombardment mass spectrometry. Lett Pept Sci 2:345–351CrossRefGoogle Scholar
  61. Perich JW, Meggio F, Pinna LA (1996b) Solid phase synthesis of pp60src-related phosphopeptides via ‘Global’ phosphorylation and their use as substrates for enzymic phosphorylation by casein kinase-2. Bioorg Med Chem 4:143–150PubMedCrossRefGoogle Scholar
  62. Perich JW, Ede NJ, Eagle S, Bray AM (1999) Synthesis of phosphopeptides by the Multipin method: evaluation of coupling methods for the incorporation of Fmoc-Tyr(PO3Bzl,H)-OH, Fmoc-Ser(PO3Bzl,H)-OH and Fmoc-Thr(PO3Bzl,H)-OH. Lett Pept Sci 6:91–97Google Scholar
  63. Poteur L, Trifilieff E (1996) Global phosphorylation at Ser16 of the 32-residue cytoplasmic domain of phospholamban: comparison of di-t-butyl- and dibenzyl-N,N-diisopropylphosphoramidites. Lett Pept Sci 2:271–276CrossRefGoogle Scholar
  64. Qabar MN, Urban J, Kahn M (1997) A facile solution and solid phase synthesis of phosphotyrosine mimetic L-4-[diethylphosphono(difluoromethyl)]phenylalanine (F2Pmp(EtO)2) derivatives. Tetrahedron Lett 53:11171–11178Google Scholar
  65. Sakaguchi K, Saito SI, Higashimoto Y, Roy S, Anderson CW, Appella E (2000) Damage-mediated phosphorylation of human p53 threonine 18 through a cascade mediated by a casein 1-like kinase. Effect on Mdm2 binding. J Biol Chem 275:9278–9283PubMedCrossRefGoogle Scholar
  66. Sakamoto H, Kodama H, Higashimoto Y, Kondo M, Lewis MS, Anderson CW, Appella E, Sakaguchi K (1996) Chemical synthesis of phosphorylated peptides of the carboxy-terminal domain of human p53 by a segment condensation method. Int J Pept Protein Res 48:429–442PubMedCrossRefGoogle Scholar
  67. Sawabe A, Filla SA, Masamune S (1992) Use of 2-(trimethylsilyl)ethyl as a protecting group in phosphate monoester synthesis. Tetrahedron Lett 33:7685–7686CrossRefGoogle Scholar
  68. Shapiro G, Buechler D, Ojea V, Pombo-Villar E, Ruiz M, Weber HP (1993) Synthesis of both D- and L-Fmoc-Abu[PO(OCH2CH:CH2)2]-OH for solid phase phosphonopeptide synthesis. Tetrahedron Lett 34:6255–6258CrossRefGoogle Scholar
  69. Shapiro G, Buechler D, Enz A, Pombo-Villar E (1994) Solid-phase synthesis of phosphonoserine isosteres of phosphoserine peptides. Tetrahedron Lett 35:1173–1176CrossRefGoogle Scholar
  70. Shen K, Keng Y-F, Wu L, Guo X-L, Lawrence DS, Zhang Z-Y (2001) Acquisition of a specific and potent PTP1B inhibitor from a novel combinatorial library and screening procedure. J Biol Chem 276:47311–47319PubMedCrossRefGoogle Scholar
  71. Solas D, Hale RL, Patel DV (1996) An efficient synthesis of N-a-Fmoc-4-(Phosphonodifluoromethyl)-L-phenylalanine. J Org Chem 61:1537–1539CrossRefGoogle Scholar
  72. Songyang Z, Shoelson SE, Chaudhuri M, Gish G, Pawson T, Haser WG, King F, Roberts T, Ratnofsky S, Lechleider RJ et al (1993) SH2 domains recognize specific phosphopeptide sequences. Cell 72:767–778PubMedCrossRefGoogle Scholar
  73. Songyang Z, Shoelson SE, Mcglade J, Olivier P, Pawson T, Bustelo XR, Barbacid M, Sabe H, Hanafusa H, Yi T et al (1994) Specific motifs recognized by the SH2 domains of Csk, 3BP2, fps/fes, GRB-2, HCP, SHC, Syk, and Vav. Mol Cell Biol 14:2777–2785PubMedGoogle Scholar
  74. Staerkaer G, Jakobsen MH, Olsen CE, Holm A (1991) Solid phase peptide synthesis of selectively phosphorylated peptides. Tetrahedron Lett 32:5389–5392CrossRefGoogle Scholar
  75. Sun J-P, Fedorov Alexander A, Lee S-Y, Guo X-L, Shen K, Lawrence David S, Almo Steven C, Zhang Z-Y (2003) Crystal structure of PTP1B complexed with a potent and selective bidentate inhibitor. J Biol Chem 278:12406–12414PubMedCrossRefGoogle Scholar
  76. Tamura K (2001) Synthesis of a phosphorylated peptide by a solid-phase method. Seirigaku Gijutsu Kenkyukai Hokoku 23:32–35Google Scholar
  77. Tegge W (1994) Solid-phase synthesis of phosphorylated and thiophosphorylated peptides related to an EGFR sequence. Int J Pept Protein Res 43:448–453PubMedCrossRefGoogle Scholar
  78. Tholey A, Reed J, Lehmann WD (1999) Electrospray tandem mass spectrometric studies of phosphopeptides and phosphopeptide analogs. J Mass Spectrom 34:117–123PubMedCrossRefGoogle Scholar
  79. Valerio RM, Bray AM, Maeji NJ, Morgan PO, Perich JW (1995) Preparation of O-phosphotyrosine-containing peptides by Fmoc solid-phase synthesis: evaluation of several Fmoc-Tyr(PO3R2)-OH derivatives. Lett Pept Sci 2:33–40CrossRefGoogle Scholar
  80. Vorherr T, Bannwarth W (1995) Phospho-serine and phospho-threonine building blocks for the synthesis of phosphorylated peptides by the Fmoc solid phase strategy. Bioorg Med Chem 5:2661–2664CrossRefGoogle Scholar
  81. Wade JD, Perich JW, Mcleish MJ, Otvos L Jr, Tregear GW (1995) Synthesis and conformational analysis of an O-phosphotyrosine-containing a-helical peptide. Lett Pept Sci 2:71–76CrossRefGoogle Scholar
  82. Wakamiya T, Saruta K, Yasuoka J, Kusumoto S (1994) An efficient procedure for solid-phase synthesis of phosphopeptides by the Fmoc strategy. Chem Lett 1099–1102Google Scholar
  83. Wakamiya T, Nishida T, Togashi R, Saruta K, Yasuoka J-I, Kusumoto S (1996) Preparations of Na-Fmoc-O-[(benzyloxy)hydroxyphosphinyl] b-hydroxy a-amino acid derivatives. Bull Chem Soc Jpn 69:465–468CrossRefGoogle Scholar
  84. Wakamiya T, Togashi R, Nishida T, Saruta K, Yasuoka J-I, Kusumoto S, Aimoto S, Kumagaye YK, Nakajima K, Nagata K (1997) Synthetic study of phosphopeptides related to heat shock protein HSP27. Bioorg Med Chem 5:135–145PubMedCrossRefGoogle Scholar
  85. Waksman G, Shoelson SE, Pant N, Cowburn D, Kuriyan J (1993) Binding of a high affinity phosphotyrosyl peptide to the Src SH2 domain: crystal structures of the complexed and peptide-free forms. Cell 72:779–790PubMedCrossRefGoogle Scholar
  86. Walsh DA, Perkins JP, Krebs EG (1968) An adenosine 3′,5′-monophosphate-dependant protein kinase from rabbit skeletal muscle. J Biol Chem 243:3763–3765PubMedGoogle Scholar
  87. White, P, Beythien J (1996) Preparation of phosphoserine, threonine and tyrosine containing peptides by the Fmoc methodology using pre-formed phosphoamino acid building blocks. Innovation and perspectives in solid phase synthesis & combinatorial libraries: peptides, proteins and nucleic acids – small molecule organic chemical diversity. Collected papers, international symposium, 4th, Edinburgh, 12–16 September 1995, pp. 557–560Google Scholar
  88. Wrobel J, Dietrich A (1993) Preparation of L-(phosphonodifluoromethyl)phenylalanine derivatives as non-hydrolyzable mimetics of O-phosphotyrosine. Tetrahedron Lett 34:3543–3546CrossRefGoogle Scholar
  89. Xu Q, Ottinger EA, Sole NA, Barany G (1997) Detection and minimization of H-phosphonate side reaction during phosphopeptide synthesis by a post-assembly global phosphorylation strategy. Lett Pept Sci 3:333–342CrossRefGoogle Scholar
  90. Zhao Z (1996) Thiophosphate derivatives as inhibitors of tyrosine phosphatases. Biochem Biophys Res Commun 218:480–484PubMedCrossRefGoogle Scholar
  91. Zheng W, Zhang Z, Ganguly S, Weller JL, Klein DC, Cole PA (2003) Cellular stabilization of the melatonin rhythm enzyme induced by nonhydrolyzable phosphonate incorporation. Nat Struct Biol 10:1054–1057PubMedCrossRefGoogle Scholar
  92. Zheng W, Schwarzer D, Lebeau A, Weller JL, Klein DC, Cole PA (2005) Cellular stability of serotonin N-Acetyltransferase conferred by phosphonodifluoromethylene alanine (Pfa) substitution for Ser-205. J Biol Chem 280:10462–10467PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Troy J. Attard
    • 1
  • Neil O’Brien-Simpson
    • 1
  • Eric C. Reynolds
    • 1
    Email author
  1. 1.Cooperative Research Centre for Oral Health Science, School of Dental Science, Bio21 Institute of Molecular Science and BiotechnologyThe University of MelbourneVICAustralia

Personalised recommendations