Optimized aminolysis conditions for cleavage of N-protected hydrophobic peptides from solid-phase resins

Authors

  • D.V. Greathouse,

  • R.L. Goforth,

  • T. Crawford,

  • P. C. A. Van Der Wel,

  • J.A. Killian


  • To cite this article:

    Greathouse, D. V., Goforth, R. L., Crawford, T., van der Wel, P. C. A. & Killian, J. A. Optimized aminolysis conditions for cleavage of N-protected hydrophobic peptides from solid-phase resins.

    J. Peptide Res. 2001, 57, 519–527.

Denise V. Greathouse
Department of Chemistry and Biochemistry
Chemistry Building, Room 101
University of Arkansas
Fayetteville
AR 72701
USA
Tel.: 1-501-575-3181
Fax: 1-501-575-4049
E-mail: dgreatho@uark.edu

Abstract

Abstract: Solid-phase synthesis and aminolysis cleavage conditions were optimized to obtain N- and C-terminally protected hydrophobic peptides with both high quality and yield. Uncharged ‘WALP’ peptides, consisting of a central (Leu-Ala)n repeating unit (where n = 5, 10.5 or 11.5) flanked on both sides by Trp ‘anchors’, and gramicidin A (gA) were synthesized using 9-fluorenylmethoxycarbonyl chemistry from either Wang or Merrifield resins. For WALP peptides, the N-terminal amino acid was capped by coupling N-acetyl- or N-formyl-Ala or -Gly to the peptide/resin or by formylation of the completed peptide/resin with para-nitrophenylformate (p-NPF). N-Terminal acetyl- or formyl-Ala racemized when coupled as an HOBt-ester to the resin-bound peptide, but not when the peptide was formylated with p-NPF. Racemization was avoided at the last step by completing the peptide with acetyl- or formyl-Gly. For both WALP peptides and gA, cleavage conditions using ethanolamine or ethylenediamine were optimized as functions of solvent, time, temperature and resin type. For WALP peptides, maximum yields of highly pure peptide were obtained by cleavage with 20% ethanolamine or ethylenediamine in 80% dichloromethane for 48 h at 24°C. N-Acetyl-protected WALP peptides consistently gave higher yields than those protected with N-formyl. For gA, cleavage with 20% ethanolamine or ethylenediamine in 80% dimethylformamide for 48 h at 24°C gave excellent results. For both WALP peptides and gA, decreasing the cleavage time to 4 h and increasing the temperature to 40–55°C resulted in significantly lower yields. The inclusion of hexafluoroisopropanol in the cleavage solvent mixture did not improve yields for either gA or WALP peptides.

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