Bulletin of the Chemical Society of Japan · 1991 · 298 citations · 7 references
Life SciencesBiochemistryMolecular BiologyProtein BiochemistryProtein EngineeringStructural BiologyAmino Acid ChemistryPeptide ChemistryProtein ChemistryProtein StructureAmino Acid ThioestersBoc ProtectionCarboxyl Protecting GroupsDna-binding MotifsPost-translationally Modified PeptidesSolid-phase Peptide SynthesisThioester Chemistry
The DNA‑binding domain of c‑Myb protein (142–193) was synthesized using this method to assess its utility. The carboxyl‑side thioester of each peptide segment was transformed into a p‑nitrophenyl ester with silver ions and p‑nitrophenol, then the two segments were condensed. The S‑alkyl thioester approach was developed, allowing purification of partially protected c‑Myb peptide segments and near‑complete coupling to yield a highly purified DNA‑binding domain.
Abstract A method for polypeptide synthesis using S-alkyl thioester of a partially protected peptide segment has been developed. The DNA-binding domain of c-Myb protein (142–193) was synthesized by this method to estimate its usefulness. The partially protected peptide segments, Boc–[Lys(Boc)143,144,160]–c-Myb protein(142–163)–SCH2CH2CONH2 and [Lys(Boc)171,182,192]–c-Myb protein(164–193)–NH2, were highly purified from peptides obtained by a solid-phase method. The thioester in the carboxyl component of the peptide segment was converted to the corresponding p-nitrophenyl ester in the presence of silver ions and p-nitrophenol and then the two segments were condensed. The coupling reaction proceeded almost completely yielding a highly-purified product after removal of Boc groups.
7
Nucleotide sequence of cDNA clones of the murine myb proto‐oncogene.
Thomas J. Gonda, N M Gough, Ashley R. Dunn et al. · The EMBO Journal · 1985 · 174 citations · Full text