Polypeptide synthesis using an expressed peptide as a building block for condensation with a peptide thioester: application to the synthesis of phosphorylated p21Max protein(1-101).
An expressed peptide proved to be useful as a building block for the synthesis of a polypeptide via the thioester method. A partially protected peptide segment, for use as a C-terminal building block, could be prepared from a recombinant protein; its N-terminal amino acid residue was transaminated to an alpha-oxoacyl group, the side-chain amino groups were then protected with t-butoxycarbonyl (Boc) groups, and. finally, the alpha-oxoacyl group was removed. On the other hand, an O-phosphoserine-containing peptide thioester was synthesized via a solid-phase method using Boc chemistry. These building blocks were then condensed in the presence of silver ions and an active ester component. During the condensation, epimerization at the condensation site could be suppressed by the use of N,N-dimthylformamide (DMF) as a solvent. Using this strategy, a phosphorylated partial peptide of the p21Max protein, [Ser(PO3H2)2.11]-p21Max(1-101), was successfully synthesized.
Kawakami
T
T
Institute for Protein Research, Osaka University, Japan.
Hasegawa
K
K
Teruya
K
K
Akaji
K
K
Horiuchi
M
M
Inagaki
F
F
Kurihara
Y
Y
Uesugi
S
S
Aimoto
S
S
D050976:Basic-Leucine Zipper Transcription Factors; D002851:Chromatography, High Pressure Liquid; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D004126:Dimethylformamide / Q000494:pharmacology; D008956:Models, Chemical; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry*; D010766:Phosphorylation; D010768:Phosphoserine / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012997:Solvents / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014157:Transcription Factors
D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; C067850:Myc associated factor X; D010455:Peptides; D012997:Solvents; D014157:Transcription Factors; D010768:Phosphoserine; D004126:Dimethylformamide