Amino Acids

, Volume 47, Issue 7, pp 1283–1299 | Cite as

Synthetic strategies for polypeptides and proteins by chemical ligation

  • Ming Chen
  • Pascal Heimer
  • Diana ImhofEmail author
Review Article


This review focuses on chemical ligation methods for the preparation of oligopeptides and proteins. Chemical ligation is a practical and convenient methodology in peptide and protein synthesis. Longer peptides and proteins can be obtained with high yield in aqueous buffer solutions by coupling unprotected peptide segments even without activation by enzymes or further chemical agents. Several methods and protocols were developed in the past. The potential of the most important approaches of the thioester- and imine-ligation techniques is demonstrated by a broad spectrum of applications. In addition, special features and protocols such as the template-directed ligation, ligation with novel additives or solvent media, microwave-assisted ligation, and the achievements obtained with those are also highlighted herein.


Chemical ligation Native chemical ligation Thioester Chemoselective synthesis Protein synthesis 



We thank the DAAD and the Chinese Scholarship Council for financial support.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Agarwal P, van der Weijden J, Sletten EM et al (2013a) A Pictet-Spengler ligation for protein chemical modification. Proc Natl Acad Sci USA 110:46–51PubMedCentralPubMedGoogle Scholar
  2. Agarwal P, Kudirka R, Albers AE et al (2013b) Hydrazino-pictet-spengler ligation as a biocompatible method for the generation of stable protein conjugates. Bioconjug Chem 24:846–851PubMedGoogle Scholar
  3. Assem N, Natarajan A, Yudin AK (2010) Chemoselective peptidomimetic ligation using thioacid peptides and aziridine templates. J Am Chem Soc 132:10986–10987PubMedGoogle Scholar
  4. Bacsa B, Kappe CO (2007) Rapid solid-phase synthesis of a calmodulin-binding peptide using controlled microwave irradiation. Nat Protoc 2:2222–2227PubMedGoogle Scholar
  5. Beekman NJCM, Schaaper WMM, Langeveld JPM et al (2001) The nature of the bond between peptide and carrier molecule determines the immunogenicity of the construct. J Pept Res 58:237–245PubMedGoogle Scholar
  6. Bodapati KC, Soudy R, Etayash H et al (2013) Design, synthesis and evaluation of antimicrobial activity of N-terminal modified Leucocin A analogues. Bioorg Med Chem 21:3715–3722PubMedGoogle Scholar
  7. Bode JW, Fox RM, Baucom KD (2006) Chemoselective amide ligations by decarboxylative condensations of N-alkylhydroxylamines and alpha-ketoacids. Angew Chem Int Ed 45:1248–1252Google Scholar
  8. Böhm M, Kühl T, Hardes K et al (2012) Synthesis and functional characterization of tridegin and its analogues: inhibitors and substrates of factor XIIIa. Chem Med Chem 7:326–333PubMedGoogle Scholar
  9. Böhm M, Tietze AA, Heimer P et al (2013) Ionic liquids as reaction media for oxidative folding and native chemical ligation of cysteine-containing peptides. J Mol Liq 192:67–70Google Scholar
  10. Boll E, Dheur J, Drobecq H, Melnyk O (2012) Access to cyclic or branched peptides using bis(2-sulfanylethyl)amido side-chain derivatives of Asp and Glu. Org Lett 14:2222–2225PubMedGoogle Scholar
  11. Boll E, Ebran J-P, Drobecq H et al (2014) Access to large cyclic peptides by a one-pot two-peptide segment ligation/cyclization process. Org Lett 17:130–133PubMedGoogle Scholar
  12. Boll E, Drobecq H, Ollivier N et al (2015) One-pot chemical synthesis of small ubiquitin-like modifier protein–peptide conjugates using bis(2-sulfanylethyl)amido peptide latent thioester surrogates. Nat Protoc 10:269–292PubMedGoogle Scholar
  13. Botti P, Pallin TD, Tam JP (1996) Cyclic peptides from linear unprotected peptide precursors through thiazolidine formation. J Am Chem Soc 118:10018–10024Google Scholar
  14. Botti P, Carrasco MR, Kent SB (2001) Native chemical ligation using removable Nα-(1-phenyl-2-mercaptoethyl) auxiliaries. Tetrahedron Lett 42:1831–1833Google Scholar
  15. Botti P, Villain M, Manganiello S, Gaertner H (2004) Native chemical ligation through in situ O to S acyl shift. Org Lett 6:4861–4864PubMedGoogle Scholar
  16. Brik A, Wong CH (2007) Sugar-assisted ligation for the synthesis of glycopeptides. Chemistry 13:5670–5675PubMedGoogle Scholar
  17. Brik A, Ficht S, Yang YY et al (2006) Sugar-assisted ligation of N-linked glycopeptides with broad sequence tolerance at the ligation junction. J Am Chem Soc 128:15026–15033PubMedGoogle Scholar
  18. Bruick RK, Dawson PE, Kent SBH et al (1996) Template-directed ligation of peptides to oligonucleotides. Chem Biol 3:49–56PubMedGoogle Scholar
  19. Burns JA, Butler JC, Moran J, Whitesides GM (1991) Selective reduction of disulfides by tris(2-carboxyethyl)phosphine. J Org Chem 56:2648–2650Google Scholar
  20. Campbell RE (2013) Synthesis of thioester peptides for traditional Native Chemical Ligation of the Syk protein and for auxiliary-mediated Native Chemical Ligation. Dissertation AAI1544248, Purdue UniversityGoogle Scholar
  21. Canne LE, Bark SJ, Kent SBH (1996) Extending the applicability of native chemical ligation. J Am Chem Soc 118:5891–5896Google Scholar
  22. Cemazar M, Craik DJ (2008) Microwave-assisted Boc-solid phase peptide synthesis of cyclic cysteine-rich peptides. J Pept Sci 14:683–689PubMedGoogle Scholar
  23. Chandrudu S, Simerska P, Toth I (2013) Chemical methods for peptide and protein production. Molecules 18:4373–4388PubMedGoogle Scholar
  24. Coin I, Beyermann M, Bienert M (2007) Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences. Nat Protoc 2:3247–3256PubMedGoogle Scholar
  25. Cowper B, Sze TM, Premdjee B et al (2015) Examination of mercaptobenzyl sulfonates as catalysts for native chemical ligation: application to the assembly of a glycosylated glucagon-like peptide 1 (GLP-1) analogue. Chem Commun 51:3208–3210Google Scholar
  26. Craik DJ (2012) Protein folding: Turbo-charged crosslinking. Nat Chem 4:600–602Google Scholar
  27. Dang B, Kubota T, Mandal K et al (2013) Native chemical ligation at Asx-Cys, Glx-Cys: chemical synthesis and high-resolution X-ray structure of ShK toxin by racemic protein crystallography. J Am Chem Soc 135:11911–11919PubMedGoogle Scholar
  28. Dawson PE, Kent SB (2000) Synthesis of native proteins by chemical ligation. Annu Rev Biochem 69:923–960PubMedGoogle Scholar
  29. Dawson P, Muir T, Clark-Lewis I, Kent S (1994) Synthesis of proteins by native chemical ligation. Science 266:776–779PubMedGoogle Scholar
  30. Dawson PE, Churchill MJ, Ghadiri MR, Kent SBH (1997) Modulation of reactivity in native chemical ligation through the use of thiol additives. J Am Chem Soc 119:4325–4329Google Scholar
  31. Diezmann F, Eberhard H, Seitz O (2010) Native chemical ligation in the synthesis of internally modified oligonucleotide-peptide conjugates. Biopolymers 94:397–404PubMedGoogle Scholar
  32. Dirksen A, Hackeng TM, Dawson PE (2006) Nucleophilic catalysis of oxime ligation. Angew Chem Int Ed Engl 45:7581–7584PubMedGoogle Scholar
  33. Dittmann M, Sadek M, Seidel R, Engelhard M (2012) Native chemical ligation in dimethylformamide can be performed chemoselectively without racemization. J Pept Sci 18:312–316PubMedGoogle Scholar
  34. Durek T, Alewood PF (2011) Preformed selenoesters enable rapid native chemical ligation at intractable sites. Angew Chem Int Ed 50:12042–12045Google Scholar
  35. Dyer FB (2013) The aziridine-mediated ligation and efforts towards a general synthesis of N-terminal aziridinyl peptides. Dissertation 3611261, Washington UniveristyGoogle Scholar
  36. Dyer FB, Park CM, Joseph R, Garner P (2011) Aziridine-mediated ligation and site-specific modification of unprotected peptides. J Am Chem Soc 133:20033–20035PubMedGoogle Scholar
  37. Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77PubMedGoogle Scholar
  38. Englebretsen DR, Garnham BC, Bergman DA, Alewood PF (1995) A novel thioether linker: chemical synthesis of a HIV-1 protease analogue by thioether ligation. Tetrahedron Lett 36:8871–8874Google Scholar
  39. Epp O, Ladenstein R, Wendel A (1983) The refined structure of the selenoenzyme glutathione peroxidase at 0.2-nm resolution. Eur J Biochem 133:51–69PubMedGoogle Scholar
  40. Erben A, Grossmann TN, Seitz O (2011) DNA-instructed acyl transfer reactions for the synthesis of bioactive peptides. Bioorg Med Chem Lett 21:4993–4997PubMedGoogle Scholar
  41. Erdélyi M, Gogoll A (2002) Rapid microwave-assisted solid phase peptide synthesis. Synthesis (Stuttg) 11:1592–1596Google Scholar
  42. Fang GM, Li YM, Shen F et al (2011) Protein chemical synthesis by ligation of peptide hydrazides. Angew Chem Int Ed 50:7645–7649Google Scholar
  43. Fang GM, Wang JX, Liu L (2012) Convergent chemical synthesis of proteins by ligation of peptide hydrazides. Angew Chem Int Ed 51:10347–10350Google Scholar
  44. Fukuda H, Irie K, Nakahara A et al (1999) Solid-phase synthesis, mass spectrometric analysis of the zinc-folding, and phorbol ester-binding studies of the 116-mer peptide containing the tandem cysteine-rich C1 domains of protein kinase C gamma. Bioorg Med Chem 7:1213–1221PubMedGoogle Scholar
  45. Gaertner HF, Rose K, Cotton R et al (1992) Construction of protein analogs by site-specific condensation of unprotected fragments. Bioconjug Chem 3:262–268PubMedGoogle Scholar
  46. Galanis AS, Albericio F, Grøtli M (2009) Solid-phase peptide synthesis in water using microwave-assisted heating. Org Lett 11:4488–4491PubMedGoogle Scholar
  47. Galonić DP, Ide ND, van der Donk WA, Gin DY (2005) Aziridine-2-carboxylic acid-containing peptides: application to solution- and solid-phase convergent site-selective peptide modification. J Am Chem Soc 127:7359–7369PubMedGoogle Scholar
  48. Galy N, Mazieres MR, Plaquevent JC (2013) Toward waste-free peptide synthesis using ionic reagents and ionic liquids as solvents. Tetrahedron Lett 54:2703–2705Google Scholar
  49. Gieselman MD, Xie L, van der Donk WA (2001) Synthesis of a selenocysteine-containing peptide by native chemical ligation. Org Lett 3:1331–1334PubMedGoogle Scholar
  50. Goldmann AS, Barner L, Kaupp M et al (2012) Orthogonal ligation to spherical polymeric microparticles: modular approaches for surface tailoring. Prog Polym Sci 37:975–984Google Scholar
  51. Greenberg ML, Cammack N (2004) Resistance to enfuvirtide, the first HIV fusion inhibitor. J Antimicrob Chemother 54:333–340PubMedGoogle Scholar
  52. Gunasekera S, Aboye TL, Madian WA et al (2013) Making ends meet: microwave-accelerated synthesis of cyclic and disulfide rich proteins via in situ thioesterification and native chemical ligation. Int J Pept Res Ther 19:43–54PubMedCentralPubMedGoogle Scholar
  53. Haase C, Rohde H, Seitz O (2008) Native chemical ligation at valine. Angew Chem Int Ed Engl 47:6807–6810PubMedGoogle Scholar
  54. Hackenberger CPR, Schwarzer D (2008) Chemoselective ligation and modification strategies for peptides and proteins. Angew Chem Int Ed 47:10030–10074Google Scholar
  55. Hackeng TM, Griffin JH, Dawson PE (1999) Protein synthesis by native chemical ligation: expanded scope by using straightforward methodology. Proc Natl Acad Sci USA 96:10068–10073PubMedCentralPubMedGoogle Scholar
  56. Hallett JP, Welton T (2011) Room-temperature ionic liquids: solvents for synthesis and catalysis. 2. Chem Rev 111:3508–3576PubMedGoogle Scholar
  57. Hansen FK, Ha K, Todadze E et al (2011) Microwave-assisted chemical ligation of S-acyl peptides containing non-terminal cysteine residues. Org Biomol Chem 9:7162–7167PubMedGoogle Scholar
  58. Heinrikson RL (1971) The selective S-methylation of sulfhydryl groups in proteins and peptides with methyl-p-nitrobenzenesulfonate. J Biol Chem 246:4090–4096PubMedGoogle Scholar
  59. Hojo H, Aimoto S (1991) Polypeptide synthesis using the S-alkyl thioester of a partially protected peptide segment. Synthesis of the DNA-binding domain of c-Myb protein (142-193)-NH2. Bull Chem Soc Jpn 64:111–117Google Scholar
  60. Hojo H, Onuma Y, Akimoto Y et al (2007) N-Alkyl cysteine-assisted thioesterification of peptides. Tetrahedron Lett 48:25–28Google Scholar
  61. Hondal RJ (2009) Using chemical approaches to study selenoproteins-focus on thioredoxin reductases. Biochim Biophys Acta 1790:1501–1512PubMedCentralPubMedGoogle Scholar
  62. Hossany BR, Johnston BD, Wen X et al (2009) Design, synthesis, and immunochemical characterization of a chimeric glycopeptide corresponding to the Shigella flexneri Y O-polysaccharide and its peptide mimic MDWNMHAA. Carbohydr Res 344:1412–1427PubMedGoogle Scholar
  63. Hou W, Zhang X, Li F, Liu CF (2011) Peptidyl N, N-bis(2-mercaptoethyl)-amides as thioester precursors for native chemical ligation. Org Lett 13:386–389PubMedGoogle Scholar
  64. Izumi M, Otsuki A, Nishihara M et al (2014) Chemical synthesis of a synthetic analogue of the sialic acid-binding lectin siglec-7. ChemBioChem 15:2503–2507PubMedGoogle Scholar
  65. Jebrail MJ, Assem N, Mudrik JM et al (2012) Combinatorial synthesis of peptidomimetics using digital microfluidics. J Flow Chem 2:103–107Google Scholar
  66. Johnson ECB, Kent SBH (2006) Insights into the mechanism and catalysis of the native chemical ligation reaction. J Am Chem Soc 128:6640–6646PubMedGoogle Scholar
  67. Johnson ECB, Kent SBH (2007) Towards the total chemical synthesis of integral membrane proteins: a general method for the synthesis of hydrophobic peptide-thioester building blocks. Tetrahedron Lett 48:1795–1799PubMedCentralPubMedGoogle Scholar
  68. Ju L, Lippert AR, Bode JW (2008) Stereoretentive synthesis and chemoselective amide-forming ligations of C-terminal peptide alpha-ketoacids. J Am Chem Soc 130:4253–4255PubMedGoogle Scholar
  69. Kawakami T, Aimoto S (2003) A photoremovable ligation auxiliary for use in polypeptide synthesis. Tetrahedron Lett 44:6059–6061Google Scholar
  70. Kern A, Seitz O (2015) Template-directed ligation on repetitive DNA sequences: a chemical method to probe the length of huntington DNA. Chem Sci 6:724–728Google Scholar
  71. Kimmerlin T, Seebach D (2005) “100 years of peptide synthesis”: ligation methods for peptide and protein synthesis with applications to beta-peptide assemblies. J Pept Res 65:229–260PubMedGoogle Scholar
  72. Kühl T, Chen M, Teichmann K et al (2014) Ionic liquid 1-ethyl-3-methylimidazolium acetate: an attractive solvent for native chemical ligation of peptides. Tetrahedron Lett 55:3658–3662Google Scholar
  73. Lescure A (1999) Novel selenoproteins identified in silico and in vivo by using a conserved RNA structural motif. J Biol Chem 274:38147–38154PubMedGoogle Scholar
  74. Lewandowski B, De Bo G, Ward JW et al (2013) Sequence-specific peptide synthesis by an artificial small-molecule machine. Science 339:189–193PubMedGoogle Scholar
  75. Li X, Zhang L, Hall SE, Tam JP (2000) A new ligation method for N-terminal tryptophan-containing peptides using the Pictet-Spengler reaction. Tetrahedron Lett 41:4069–4073Google Scholar
  76. Li J, Cui HK, Liu L (2010) Peptide ligation assisted by an auxiliary attached to amidyl nitrogen. Tetrahedron Lett 51:1793–1796Google Scholar
  77. Lidström P, Tierney J, Wathey B, Westman J (2001) Microwave assisted organic synthesis: a review. Tetrahedron 57:9225–9283Google Scholar
  78. Liu CF, Tam JP (1994a) Peptide segment ligation strategy without use of protecting groups. Proc Natl Acad Sci 91:6584–6588PubMedCentralPubMedGoogle Scholar
  79. Liu CF, Tam JP (1994b) Chemical ligation approach to form a peptide bond between unprotected peptide segments: concept and model study. J Am Chem Soc 116:4149–4153Google Scholar
  80. Liu CF, Rao C, Tam JP (1996a) Acyl disulfide-mediated intramolecular acylation for orthogonal coupling between unprotected peptide segments: mechanism and application. Tetrahedron Lett 37:933–936Google Scholar
  81. Liu CF, Rao C, Tam JP (1996b) Orthogonal ligation of unprotected peptide segments through pseudoproline formation for the synthesis of HIV-1 protease analogs. J Am Chem Soc 118:307–312Google Scholar
  82. Liu Y, Sha R, Wang R et al (2008) 2′,2′-Ligation demonstrates the thermal dependence of DNA-directed positional control. Tetrahedron 64:8417–8422PubMedCentralPubMedGoogle Scholar
  83. Low SC, Harney JW, Berry MJ (1995) Cloning and functional characterization of human selenophosphate synthetase, an essential component of selenoprotein synthesis. J Biol Chem 270:21659–21664PubMedGoogle Scholar
  84. Low DW, Hill MG, Carrasco MR et al (2001) Total synthesis of cytochrome b562 by native chemical ligation using a removable auxiliary. Proc Natl Acad Sci USA 98:6554–6559PubMedCentralPubMedGoogle Scholar
  85. Maki T, Kawamura A, Kato N, Ohkanda J (2013) Chemical ligation of epoxide-containing fusicoccins and peptide fragments guided by 14-3-3 protein. Mol Bio Syst 9:940–943Google Scholar
  86. Malins LR, Payne RJ (2012) Synthesis and utility of β-selenol-phenylalanine for native chemical ligation-deselenization chemistry. Org Lett 14:3142–3145PubMedGoogle Scholar
  87. Malins LR, Payne RJ (2014) Recent extensions to native chemical ligation for the chemical synthesis of peptides and proteins. Curr Opin Chem Biol 22C:70–78Google Scholar
  88. Malins LR, Mitchell NJ, Payne RJ (2014) Peptide ligation chemistry at selenol amino acids. J Pept Sci 20:64–77PubMedGoogle Scholar
  89. Marinzi C, Offer J, Longhi R, Dawson PE (2004) An o-nitrobenzyl scaffold for peptide ligation: synthesis and applications. Bioorg Med Chem 12:2749–2757PubMedGoogle Scholar
  90. Medini K, Harris PWR, Hards K et al (2015) Chemical synthesis of a pore-forming antimicrobial protein, caenopore-5, by using native chemical ligation at a Glu-Cys site. Chem Bio Chem 16:328–336PubMedGoogle Scholar
  91. Melnyk O, Agouridas V (2014) From protein total synthesis to peptide transamidation and metathesis: playing with the reversibility of N, S-acyl or N, Se-acyl migration reactions. Curr Opin Chem Biol 22:137–145PubMedGoogle Scholar
  92. Metanis N, Hilvert D (2012) Strategic use of non-native diselenide bridges to steer oxidative protein folding. Angew Chem Int Ed Engl 51:5585–5588PubMedGoogle Scholar
  93. Metanis N, Keinan E, Dawson PE (2010) Traceless ligation of cysteine peptides using selective deselenization. Angew Chem Int Ed 49:7049–7053Google Scholar
  94. Monsó M, Kowalczyk W, Andreu D, de la Torre BG (2012) Reverse thioether ligation route to multimeric peptide antigens. Org Biomol Chem 10:3116–3121PubMedGoogle Scholar
  95. Moroder L (2005) Isosteric replacement of sulfur with other chalcogens in peptides and proteins. J Pept Sci 11:187–214PubMedGoogle Scholar
  96. Moyal T, Hemantha HP, Siman P et al (2013) Highly efficient one-pot ligation and desulfurization. Chem Sci 4:2496Google Scholar
  97. Mustacich D, Powis G (2000) Thioredoxin reductase. Biochem J 346:1PubMedCentralPubMedGoogle Scholar
  98. Naider FR, Becker JM (1997) Synthesis of prenylated peptides and peptide esters. Pept Sci 43:3–14Google Scholar
  99. Nakamura T, Shigenaga A, Sato K et al (2014) Examination of native chemical ligation using peptidyl prolyl thioester. Chem Commun 50:58–60Google Scholar
  100. Offer J (2010) Native chemical ligation with Nalpha acyl transfer auxiliaries. Biopolymers 94:530–541PubMedGoogle Scholar
  101. Offer J, Dawson PE (2000) Nalpha-2-mercaptobenzylamine-assisted chemical ligation. Org Lett 2:23–26PubMedGoogle Scholar
  102. Offer J, Boddy CNC, Dawson PE (2002) Extending synthetic access to proteins with a removable acyl transfer auxiliary. J Am Chem Soc 124:4642–4646PubMedGoogle Scholar
  103. Offord RE (1969) Protection of peptides of biological origin for use as intermediates in the chemical synthesis of proteins. Nature 221:37–40PubMedGoogle Scholar
  104. Ogunkoya AO, Pattabiraman VR, Bode JW (2012) Sequential ??-ketoacid-hydroxylamine (KAHA) ligations: synthesis of C-terminal variants of the modifier protein UFM1. Angew Chem Int Ed 51:9693–9697Google Scholar
  105. Ollivier N, Dheur J, Mhidia R et al (2010) Bis(2-sulfanylethyl)amino native peptide ligation. Org Lett 12:5238–5241PubMedGoogle Scholar
  106. Ollivier N, Vicogne J, Vallin A et al (2012) A one-pot three-segment ligation strategy for protein chemical synthesis. Angew Chem Int Ed 51:209–213Google Scholar
  107. Ollivier N, Blanpain A, Boll E et al (2014) Selenopeptide transamidation and metathesis. Org Lett 16:4032–4035PubMedGoogle Scholar
  108. Olschewski D, Becker CFW (2008) Chemical synthesis and semisynthesis of membrane proteins. Mol BioSyst 4:733–740PubMedGoogle Scholar
  109. Palasek SA, Cox ZJ, Collins JM (2007) Limiting racemization and aspartimide formation in microwave-enhanced Fmoc solid phase peptide synthesis. J Pept Sci 13:143–148PubMedGoogle Scholar
  110. Pattabiraman VR, Ogunkoya AO, Bode JW (2012) Chemical protein synthesis by chemoselective alpha-ketoacid-hydroxylamine (KAHA) ligations with 5-oxaproline. Angew Chemie Int Ed 51:5114–5118Google Scholar
  111. Payne RJ, Ficht S, Tang S et al (2007) Extended sugar-assisted glycopeptide ligations: development, scope, and applications. J Am Chem Soc 129:13527–13536PubMedGoogle Scholar
  112. Quaderer R, Sewing A, Hilvert D (2001) Selenocysteine-mediated native chemical ligation. Helv Chim Acta 84:1197–1206Google Scholar
  113. Raibaut L, Ollivier N, Melnyk O (2012) Sequential native peptide ligation strategies for total chemical protein synthesis. Chem Soc Rev 41:7001–7015PubMedGoogle Scholar
  114. Raibaut L, Vicogne J, Leclercq B et al (2013) Total synthesis of biotinylated N domain of human hepatocyte growth factor. Bioorg Med Chem 21:3486–3494PubMedGoogle Scholar
  115. Rasale DB, Maity I, Das AK (2014) In situ generation of redox active peptides driven by selenoester mediated native chemical ligation. Chem Commun 50:11397–11400Google Scholar
  116. Rashidian M, Mahmoodi MM, Shah R et al (2013) A highly efficient catalyst for oxime ligation and hydrazone-oxime exchange suitable for bioconjugation. Bioconjug Chem 24:333–342PubMedCentralPubMedGoogle Scholar
  117. Ratnaparkhi MP, Chaudhari SP, Pandya VA (2011) Peptides and proteins in pharmaceuticals. Int J Curr Pharm Res 3:1–9Google Scholar
  118. Reif A, Siebenhaar S, Tröster A et al (2014) Semisynthesis of biologically active glycoforms of the human cytokine interleukin 6. Angew Chem Int Ed 53:12125–12131Google Scholar
  119. Robey FA, Fields RL (1989) Automated synthesis of N-bromoacetyl-modified peptides for the preparation of synthetic peptide polymers, peptide-protein conjugates, and cyclic peptides. Anal Biochem 177:373–377PubMedGoogle Scholar
  120. Rohde H, Schmalisch J, Harpaz Z et al (2011) Ascorbate as an alternative to thiol additives in native chemical ligation. ChemBioChem 12:1396–1400PubMedGoogle Scholar
  121. Roloff A, Seitz O (2013a) The role of reactivity in DNA templated native chemical PNA ligation during PCR. Bioorg Med Chem 21:3458–3464PubMedGoogle Scholar
  122. Roloff A, Seitz O (2013b) Bioorthogonal reactions challenged: DNA templated native chemical ligation during PCR. Chem Sci 4:432Google Scholar
  123. Rose K (1994) Facile synthesis of homogeneous artificial proteins. J Am Chem Soc 116:30–33Google Scholar
  124. Ruff Y, Garavini V, Giuseppone N (2014) Reversible native chemical ligation: a facile access to dynamic covalent peptides. J Am Chem Soc 136:6333–6339PubMedGoogle Scholar
  125. Santhakumar G, Payne RJ (2014) Total synthesis of polydiscamides B, C, and D via a convergent native chemical ligation–oxidation strategy. Org Lett 16:4500–4503PubMedGoogle Scholar
  126. Sato K, Shigenaga A, Tsuji K et al (2011) N-sulfanylethylanilide peptide as a crypto-thioester peptide. Chem Bio Chem 12:1840–1844PubMedGoogle Scholar
  127. Schnolzer M, Kent S (1992) Constructing proteins by dovetailing unprotected synthetic peptides: backbone-engineered HIV protease. Science 256:221–225PubMedGoogle Scholar
  128. Selvasekaran J, Turnbull KD (1999) Chemical ligation of oligodeoxyribonucleotides on circular DNA templates. Nucleic Acids Res 27:624–627PubMedCentralPubMedGoogle Scholar
  129. Shao J, Tam JP (1995) Unprotected peptides as building blocks for the synthesis of peptide dendrimers with oxime, hydrazone, and thiazolidine linkages. J Am Chem Soc 117:3893–3899Google Scholar
  130. Shao Y, Lu W, Kent SBH (1998) A novel method to synthesize cyclic peptides. Tetrahedron Lett 39:3911–3914Google Scholar
  131. Siman P, Karthikeyan SV, Brik A (2012) Native chemical ligation at glutamine. Org Lett 14:1520–1523PubMedGoogle Scholar
  132. Sohma Y, Kent SBH (2009) Biomimetic synthesis of lispro insulin via a chemically synthesized “mini-proinsulin” prepared by oxime-forming ligation. J Am Chem Soc 131:16313–16318PubMedCentralPubMedGoogle Scholar
  133. Sohma Y, Kitamura H, Kawashima H et al (2011) Synthesis of an O-acyl isopeptide by using native chemical ligation to efficiently construct a hydrophobic polypeptide. Tetrahedron Lett 52:7146–7148Google Scholar
  134. Stanchev S, Zawada Z, Monincová L et al (2014) Synthesis of lucifensin by native chemical ligation and characteristics of its isomer having different disulfide bridge pattern. J Pept Sci 20:725–735PubMedGoogle Scholar
  135. Steinhagen M, Zunker K, Nordsieck K, Beck-Sickinger AG (2013) Large scale modification of biomolecules using immobilized sortase A from Staphylococcus aureus. Bioorg Med Chem 21:3504–3510PubMedGoogle Scholar
  136. Tam JP, Miao Z (1999) Stereospecific pseudoproline ligation of n-terminal serine, threonine, or cysteine-containing unprotected peptides. J Am Chem Soc 121:9013–9022Google Scholar
  137. Tam JP, Yu Q (1998) Methionine ligation strategy in the biomimetic synthesis of parathyroid hormones. Biopolymers 46:319–327PubMedGoogle Scholar
  138. Tam JP, Lu YA, Liu CF, Shao J (1995) Peptide synthesis using unprotected peptides through orthogonal coupling methods. Proc Natl Acad Sci 92:12485–12489PubMedCentralPubMedGoogle Scholar
  139. Tam JP, Yu Q, Miao Z (1999) Orthogonal ligation strategies for peptide and protein. Biopolymers 51:311–332PubMedGoogle Scholar
  140. Tam JP, Xu J, Eom KD (2001) Methods and strategies of peptide ligation. Biopolymers 60:194–205PubMedGoogle Scholar
  141. Tan AR, Swain SM (2003) Ongoing adjuvant trials with trastuzumab in breast cancer. Semin Oncol 30(5 Suppl 16):54–64PubMedGoogle Scholar
  142. Tanaka S, Moriwaki S, Uenishi K et al (2011) The availability of urinary gamma-glutamyltransferase as a screening for osteoporosis. Bone 48:S210Google Scholar
  143. Tanaka T, Wagner AM, Warner JB et al (2013) Expressed protein ligation at methionine: N-terminal attachment of homocysteine, ligation, and masking. Angew Chem Int Ed Engl 52:6210–6213PubMedGoogle Scholar
  144. Thapa P, Zhang RY, Menon V, Bingham JP (2014) Native chemical ligation: a boon to peptide chemistry. Molecules 19:14461–14483PubMedGoogle Scholar
  145. Thompson RE, Liu X, Alonso-García N et al (2014) Trifluoroethanethiol: an additive for efficient one-pot peptide ligation: desulfurization chemistry. J Am Chem Soc 136:8161–8164PubMedGoogle Scholar
  146. Tolbert TJ, Franke D, Wong C-H (2005) A new strategy for glycoprotein synthesis: ligation of synthetic glycopeptides with truncated proteins expressed in E. coli as TEV protease cleavable fusion protein. Bioorg Med Chem 13:909–915PubMedGoogle Scholar
  147. Tong X, Xiao XH, Deng JC et al (2010) Applications of low temperature microwave technique in chemistry research. Prog Chem 22:2462–2468Google Scholar
  148. Tuchscherer G (1993) Template assembled synthetic proteins: condensation of a multifunctional peptide to a topological template via chemoselective ligation. Tetrahedron Lett 34:8419–8422Google Scholar
  149. Van de Langemheen H, Quarles van Ufford HLC, Kruijtzer JAW, Liskamp RMJ (2014a) Efficient synthesis of protein mimics by sequential native chemical ligation. Org Lett 16:2138–2141PubMedGoogle Scholar
  150. Van de Langemheen H, van Hoeke M, Quarles van Ufford HC et al (2014b) Scaffolded multiple cyclic peptide libraries for protein mimics by native chemical ligation. Org Biomol Chem 12:4471–4478PubMedGoogle Scholar
  151. Van de Vijver P, Scheer L, van Beijnum J et al (2012) Application of an omonasteine ligation strategy for the total chemical synthesis of the BRD7 bromodomain. Chem Commun (Camb) 48:9403–9405Google Scholar
  152. Vázquez O, Seitz O (2014) Templated native chemical ligation: peptide chemistry beyond protein synthesis. J Pept Sci 20:78–86PubMedGoogle Scholar
  153. Villain M, Gaertner H, Botti P (2003) Native chemical ligation with aspartic and glutamic acids as C-terminal residues: scope and limitations. Eur J Org Chem 2003(17):3267–3272Google Scholar
  154. Vlieghe P, Lisowski V, Martinez J, Khrestchatisky M (2010) Synthetic therapeutic peptides: science and market. Drug Discov Today 15:40–56PubMedGoogle Scholar
  155. Wagner M, Sonntag D, Grimm R et al (1999) Substrate-specific selenoprotein B of glycine reductase from Eubacterium acidaminophilum: biochemical and molecular analysis. Eur J Biochem 260:38–49PubMedGoogle Scholar
  156. Wan Q, Danishefsky SJ (2007) Free-radical-based, specific desulfurization of cysteine: a powerful advance in the synthesis of polypeptides and glycopolypeptides. Angew Chem Int Ed Engl 46:9248–9252PubMedGoogle Scholar
  157. Wang JX, Fang GM, He Y et al (2015) Peptide o-aminoanilides as crypto-thioesters for protein chemical synthesis. Angew Chem Int Ed Engl 54:2194–2198PubMedGoogle Scholar
  158. Warren JD, Miller JS, Keding SJ, Danishefsky SJ (2004) Toward fully synthetic glycoproteins by ultimately convergent routes: a solution to a long-standing problem. J Am Chem Soc 126:6576–6578PubMedGoogle Scholar
  159. Wermeling DP (2005) Ziconotide, an intrathecally administered N-type calcium channel antagonist for the treatment of chronic pain. Pharmacotherapy 25:1084–1094PubMedGoogle Scholar
  160. Wessjohann LA, Schneider A, Abbas M, Brandt W (2007) Selenium in chemistry and biochemistry in comparison to sulfur. Biol Chem 388:997–1006PubMedGoogle Scholar
  161. White P, Keyte JW, Bailey K, Bloomberg G (2004) Expediting the Fmoc solid phase synthesis of long peptides through the application of dimethyloxazolidine dipeptides. J Pept Sci 10:18–26PubMedGoogle Scholar
  162. Wieland T, Bokelmann E, Bauer L et al (1953) Über Peptidsynthesen. 8. Mitteilung Bildung von S-haltigen Peptiden durch intramolekulare Wanderung von Aminoacylresten. Justus Liebigs Ann Chem 583:129–149Google Scholar
  163. Wilce JA, Love SG, Richardson SJ et al (2001) Synthesis of an analog of the thyroid hormone-binding protein transthyretin via regioselective chemical ligation. J Biol Chem 276:25997–26003PubMedGoogle Scholar
  164. Wu J, Ruiz-Rodríguez J, Comstock JM et al (2011) Synthesis of human GLP-1 (7–36) by chemoselective α-ketoacid–hydroxylamine peptide ligation of unprotected fragments. Chem Sci 2:1976Google Scholar
  165. Yan LZ, Dawson PE (2001) Synthesis of peptides and proteins without cysteine residues by native chemical ligation combined with desulfurization. J Am Chem Soc 123:526–533PubMedGoogle Scholar
  166. Yang YY, Ficht S, Brik A, Wong CH (2007) Sugar-assisted ligation in glycoprotein synthesis. J Am Chem Soc 129:7690–7701PubMedCentralPubMedGoogle Scholar
  167. Yang R, Hou W, Zhang X, Liu CF (2012) N-to-C sequential ligation using peptidyl N, N -bis(2-mercaptoethyl)amide building blocks. Org Lett 14:374–377PubMedGoogle Scholar
  168. Yang R, Bi X, Li F et al (2014) Native chemical ubiquitination using a genetically incorporated azidonorleucine. Chem Commun 50:7971–7974Google Scholar
  169. Yu HM, Chen ST, Wang KT (1992) Enhanced coupling efficiency in solid-phase peptide synthesis by microwave irradiation. J Org Chem 57:4781–4784Google Scholar
  170. Zhang L, Tam JP (1997) Orthogonal coupling of unprotected peptide segments through histidyl amino terminus. Tetrahedron Lett 38:3–6Google Scholar
  171. Zhang L, Torgerson TR, Liu XY et al (1998) Preparation of functionally active cell-permeable peptides by single-step ligation of two peptide modules. Proc Natl Acad Sci USA 95:9184–9189PubMedCentralPubMedGoogle Scholar
  172. Zhang C, Li Y, Zhang M, Li X (2012) DNA-directed formation of peptide bond: a model study toward DNA-programmed peptide ligation. Tetrahedron 68:5152–5156Google Scholar
  173. Zheng JS, Chang HN, Wang FL, Liu L (2011) Fmoc synthesis of peptide thioesters without post-chain-assembly manipulation. J Am Chem Soc 133:11080–11083PubMedGoogle Scholar
  174. Zheng JS, Tang S, Qi YK et al (2013) Chemical synthesis of proteins using peptide hydrazides as thioester surrogates. Nat Protoc 8:2483–2495PubMedGoogle Scholar
  175. Zhong W, Skwarczynski M, Fujita Y et al (2009) Design and synthesis of lipopeptide–carbohydrate assembled multivalent vaccine candidates using native chemical ligation. Aust J Chem 62:993Google Scholar

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© Springer-Verlag Wien 2015

Authors and Affiliations

  1. 1.Pharmaceutical Chemistry I, Institute of PharmacyUniversity of BonnBonnGermany

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