Protocol abstract
Nature Protocols 2, - 3247 - 3256 (2007)
Published online: 13 December 2007 | doi:10.1038/nprot.2007.454
Subject Categories: Biochemistry and protein analysis | Synthetic chemistry
Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences
Irene Coin1, Michael Beyermann1 & Michael Bienert1
Abstract
This protocol for solid-phase peptide synthesis (SPPS) is based on the widely used Fmoc/tBu strategy, activation of the carboxyl groups by aminium-derived coupling reagents and use of PEG-modified polystyrene resins. A standard protocol is described, which was successfully applied in our lab for the synthesis of the corticotropin-releasing factor (CRF), >400 CRF analogs and a countless number of other peptides. The 41-mer peptide CRF is obtained within 
80 working hours. To achieve the so-called difficult sequences, special techniques have to be applied in order to reduce aggregation of the growing peptide chain, which is the main cause of failure for peptide chemosynthesis. Exemplary application of depsipeptide and pseudoproline units is shown for synthesizing an extremely difficult sequence, the Asn(15) analog of the WW domain FBP28, which is impossible to obtain using the standard protocol.
- Leibniz-Institut für Molekulare Pharmakologie, Department of Peptide Chemistry and Biochemistry, Robert-Rössle-Strasse 10, Berlin 13125, Germany.
Correspondence to: Irene Coin1 e-mail: coin@fmp-berlin.de
Correspondence to: Michael Beyermann1 e-mail: beyermann@fmp-berlin.de
nature-products
A-Z product listing
- 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo-[4,5-b] pyridinium hexafluorophosphate 3-oxide(GL Biochem)
- 1-[Bis(dimethylamino)methylene]-1H-benzotriazolium hexafluorophosphate 3-oxide(Iris Biotech GmbH)
- 1-Hydroxybenzotriazole(Fluka)
- Acetic anhydride(Fluka)
- Bsmoc-Xxx-OH(Morre-Tec Ind. Inc)
- Diethyl ether(Acros)
