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Volumn 129, Issue 24, 2007, Pages 7690-7701

Sugar-assisted ligation in glycoprotein synthesis

Author keywords

[No Author keywords available]

Indexed keywords

ACETAMIDOMETHYL; GLYCOPEPTIDES; LIGATION; NATIVE CHEMICAL LIGATION (NCL);

EID: 34250850156     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja0708971     Document Type: Article
Times cited : (104)

References (63)
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    • (a) Sears, P. Science 2001, 291, 2344-2350.
    • (2001) Science , vol.291 , pp. 2344-2350
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    • 33748483846 scopus 로고    scopus 로고
    • (c) Hamilton, S. R., et al. Science 2006, 313, 1441-1443.
    • (2006) Science , vol.313 , pp. 1441-1443
    • Hamilton, S.R.1
  • 9
    • 32344449790 scopus 로고    scopus 로고
    • (d) Li, H., et al. Nat. Biotechnol. 2006, 24, 210-215.
    • (2006) Nat. Biotechnol , vol.24 , pp. 210-215
    • Li, H.1
  • 10
    • 0036462604 scopus 로고    scopus 로고
    • (a) Davis, B. G. Chem. Rev. 2002, 102, 579-602.
    • (2002) Chem. Rev , vol.102 , pp. 579-602
    • Davis, B.G.1
  • 37
    • 34250805540 scopus 로고    scopus 로고
    • Molecular simulations of transition states have been performed and will be presented in a following publication. The simulations support the hypothesis that the restricted conformation of the sugar helps to preorganize the GalNAc-linked thioester intermediate and N-terminal amine for nucleophilic attack
    • Molecular simulations of transition states have been performed and will be presented in a following publication. The simulations support the hypothesis that the restricted conformation of the sugar helps to preorganize the GalNAc-linked thioester intermediate and N-terminal amine for nucleophilic attack.
  • 51
    • 34250817646 scopus 로고    scopus 로고
    • 1/2(h) for the Gly-Val junction for which the ligation efficiency is more susceptible to varying the reaction conditions: 18 h for N-linked glycopeptides (6 M Gn·HCl, 0.1 M PBS, 2% PhSH, glycopeptide (1.2 equiv), peptide thioester (1 equiv)); 14 h for α-O-linked glycopeptides (6 M Gn·HCl, 0.1 M PBS, glycopeptide (1 equiv), peptide thioester (1.2 equiv)).
    • 1/2(h) for the Gly-Val junction for which the ligation efficiency is more susceptible to varying the reaction conditions: 18 h for N-linked glycopeptides (6 M Gn·HCl, 0.1 M PBS, 2% PhSH, glycopeptide (1.2 equiv), peptide thioester (1 equiv)); 14 h for α-O-linked glycopeptides (6 M Gn·HCl, 0.1 M PBS, glycopeptide (1 equiv), peptide thioester (1.2 equiv)).
  • 52
    • 34250799266 scopus 로고    scopus 로고
    • The initial pH of the buffer used for SAL is 8.5. After addition of the peptides, the pH drops due to the residual TFA present in HPLC purified peptides. As a consequence, in some cases, triethanolamine was added to adjust the reaction pH to the ideal range 7.2-7.4.
    • The initial pH of the buffer used for SAL is 8.5. After addition of the peptides, the pH drops due to the residual TFA present in HPLC purified peptides. As a consequence, in some cases, triethanolamine was added to adjust the reaction pH to the ideal range 7.2-7.4.
  • 53
    • 34250859048 scopus 로고    scopus 로고
    • The glycopeptide was consumed by two reaction paths: reacting with the peptide thioester Cys(Acm)37-Gly52 and by generating a degraded fragment. The mass of this degraded fragment (observed mass, 3901 Da) corresponds to a loss of the first two N-terminal residues of glycopeptide Val53-Phe82. The same fragment mass was also observed in the cases of glycopeptide His53-Phe82 and glycopeptide Gly53-Phe82 by LC/MS. The first observation of this degraded fragment was in the crude glycopeptide cleaved from the resin. In the beginning of the ligation, the amount of this degraded fragment is less than 5, However, its amount increased during the course of the reaction. A similar degradation phenomenon was not observed for the short glycopeptides Table 1
    • 82 by LC/MS. The first observation of this degraded fragment was in the crude glycopeptide cleaved from the resin. In the beginning of the ligation, the amount of this degraded fragment is less than 5%. However, its amount increased during the course of the reaction. A similar degradation phenomenon was not observed for the short glycopeptides (Table 1).
  • 56
    • 0041783935 scopus 로고    scopus 로고
    • It caught our attention that Asp or Glu on the C-terminus of the peptide thioester has the potential to form an intramolecular anhydride during NCL, which could lead to the generation of a branched ligation product. To avoid this problem, the original Asp36 of diptericin was mutated to Asn 36. Alternatively, an orthogonal protecting group could be used to avoid this side reaction Villain, M, Gaertner, H, Botti, P, Chem. 2003, 3267-3272
    • 36. Alternatively, an orthogonal protecting group could be used to avoid this side reaction (Villain, M.; Gaertner, H.; Botti, P. Eur. J. Org. Chem. 2003, 3267-3272).
  • 60
    • 34250804949 scopus 로고    scopus 로고
    • tBu)-(Hmb)Gly-OH in SPPS failed due to the difficulty of complete removal of the Hmb group in the final global deprotection step.
    • tBu)-(Hmb)Gly-OH in SPPS failed due to the difficulty of complete removal of the Hmb group in the final global deprotection step.
  • 62
    • 33847791752 scopus 로고    scopus 로고
    • While we were preparing this manuscript, Kent's group reported selective desulfurization of cysteine in peptides containing Cys(Acm) residues (Pentelute, B. L, Kent, S. B. H. Org. Lett. 2007, 9, 687-690, In their report, Raney nickel was used as the catalyst. In our studies, we found that the thiol handle of the sugar moiety reacts much more rapidly towards desulfurization than the cysteine residue (data not shown, when Pd/Al 2O3 was used as the catalyst. This indicates that even better desulfurization selectivity might be achieved if different catalysts, like Raney nickel, or other conditions were used to selectively desulfurize the thiol handle in the presence of Cys(Acm) residues
    • 3 was used as the catalyst. This indicates that even better desulfurization selectivity might be achieved if different catalysts, like Raney nickel, or other conditions were used to selectively desulfurize the thiol handle in the presence of Cys(Acm) residues.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.