Incomplete Fmoc deprotection in solid-phase synthesis of peptides.
During solid-phase peptide synthesis of homo-oligopeptides containing leucine or alanine using the Fmoc strategy, we have observed ineffective N-alpha-deprotection with piperidine in a sequence-dependent manner. Incomplete deprotection was found to be associated with subsequent slow or incomplete amino acid coupling. Optimization of the deprotection step was carried out by varying the experimental conditions e.g. deprotection time, temperature, solvents and addition of chaotropes. Coupling and deprotection steps have been investigated using color monitoring, as well as FAB MS and HPLC for product analysis. The phenomena of difficult coupling and deprotection steps in the investigated systems have been demonstrated to have the same physical chemical origins, beta-sheet formation.
Larsen
B D
BD
Department of Chemical Physics, H. C. Orsted Institute, University of Copenhagen, Denmark.
Holm
A
A
D000215:Acylation; D000595:Amino Acid Sequence; D000596:Amino Acids; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D005449:Fluorenes; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry; D017433:Protein Structure, Secondary; D016339:Spectrometry, Mass, Fast Atom Bombardment; D014269:Trifluoroacetic Acid
D000596:Amino Acids; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D009842:Oligopeptides; D014269:Trifluoroacetic Acid