𝔖 Bobbio Scriptorium
✦   LIBER   ✦

A Method for Selective Isolation of the Amino-Terminal Peptide from α-Amino-Blocked Proteins

✍ Scribed by T.H. Akiyama; T. Sasagawa; M. Suzuki; K. Titani

Publisher
Elsevier Science
Year
1994
Tongue
English
Weight
377 KB
Volume
222
Category
Article
ISSN
0003-2697

No coin nor oath required. For personal study only.

✦ Synopsis

A method for selective isolation of the amino (\mathrm{N})-terminal peptide from an (\alpha)-amino (\left(N^{\alpha}\right.) )-blocked protein is presented. The method consists of four steps. First, (\epsilon-) amino groups of lysine residues in the protein are succinylated. Second, the derivatized protein is digested by either enzymatic or chemical cleavage. Third, the digest is subjected to reaction with cyanogen bromide-activated Sepharose. Only the N-terminal-blocked peptide fails to react, while the other peptides are covalently bound to the Sepharose. Fourth, uncoupled peptides are purified by reversed-phase chromatography. The amino acid sequence of the peptide including the blocking group can be determined by mass spectrometry. These procedures were successfully tested with three known (\mathbf{N}^{\alpha})-blocked proteins including bovine brain ($ 100) protein, horse cytochrome (c), and ovalbumin. a 1994 Academic Press, Inc.

📜 SIMILAR VOLUMES

Selective isolation of free and blocked
Selective isolation of free and blocked amino-terminal peptides from enzymatic digestion of proteins
✍ Harvey Kaplan; George Oda 📂 Article 📅 1983 🏛 Elsevier Science 🌐 English ⚖ 425 KB

A general method for the selective isolation of free and blocked amino-terminal peptides from proteins is described. The rationale behind the methodology is based on the reasoning that if a protein, which has all its free amino groups blocked by citraconylation, is digested with a protease, all pept

A Method for Identifying the Carboxy Ter
A Method for Identifying the Carboxy Terminal Amino Acid of a Protein
✍ T. Hayashi; T. Sasagawa 📂 Article 📅 1993 🏛 Elsevier Science 🌐 English ⚖ 437 KB

A highly sensitive new method for identifying the carboxy terminus of a protein was developed. The carboxyl terminal amino acid was racemized by reaction with acetic anhydride. The resulting modified protein was subjected to acid hydrolysis. The hydrolysate was derivatized with (+)-1-(9-fluorenyl)et

A Method for Preparation of Amino Acid T
A Method for Preparation of Amino Acid Thiohydantoins from Free Amino Acids Activated by Acetyl Chloride for Development of Protein C-Terminal Sequencing
✍ Bilan Mo; Jiang Li; Songping Liang 📂 Article 📅 1997 🏛 Elsevier Science 🌐 English ⚖ 109 KB

A novel and efficient method to prepare amino acid thiohydantoins, which are required as reference standards for development of C-terminal protein sequencing, is reported. Amino acid thiohydantoins were prepared using a straightforward method involving reaction of 20 free amino acids with acetyl chl