Chemoselective one-step purification method for peptides synthesized by the solid-phase technique.
AUTOR(ES)
Funakoshi, S
RESUMO
The specific reaction between SH and iodoacetamide groups has been explored as the basis of an affinity-type purification procedure for peptides synthesized by the solid-phase technique. For this affinity-type purification procedure, we synthesized an SH precursor reagent bearing an acid-labile S-protecting group, pMB-SCH2CONHCH2CH2-SO2CH2CH2OCO2pNP (compound I), in which pMB is p-methoxybenzyl and pNP is p-nitrophenyl. Using this reagent, the procedure involves the following sequence of four reactions: (i) attachment of the SH function of compound I to the alpha-amino group of a peptide-resin through a base-labile sulfonylethoxycarbonyl linkage in the final step of solid-phase peptide synthesis, (ii) acid treatment to remove the S-pMB and side-chain-protecting groups employed and cleave the modified peptide from the resin, (iii) immobilization of the derived SH-peptide on an iodoacetamide-resin column, and (iv) base (5% NH4OH) treatment to release the desired peptide from the resin in nearly pure form. To facilitate this purification procedure, unreacted amino groups were acetylated in each step during solid-phase synthesis. The usefulness of this method was demonstrated by the purification of several peptides (18 to approximately 44 amino acids in length) synthesized by the 9-fluorenylmethoxycarbonyl (Fmoc)-based solid-phase technique. The principle of this affinity-type purification procedure may also be applied to the tert-butoxycarbonyl (Boc)-based solid-phase technique.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=52217Documentos Relacionados
- Specificity control in solid-phase enzyme immunoassay for HBsAg by one-step in situ blocking with human anti-HBs.
- One-step affinity purification protocol for human telomerase.
- One-step purification of nisin A by immunoaffinity chromatography.
- Pyrosequencing™ : A one-step method for high resolution HLA typing
- A one-step method for in vitro production of tRNA transcripts