Development of a method for protein labeling by the use of intramolecular thioester-forming unit

2014 Fiscal Year Final Research Report

• Project/Area Number: 24550187
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Chemistry related to living body
• Research Institution: Osaka University
• Principal Investigator: KAWAKAMI Toru 大阪大学, たんぱく質研究所, 准教授 (70273711)
• Co-Investigator(Renkei-kenkyūsha): HIRAKI Yuji 京都大学, 再生医科学研究所, 教授 (40144498)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 生体分子 / ペプチド / タンパク質 / 受容体 / チオエステル / 部位特異的修飾

Outline of Final Research Achievements

We had found that a peptide containing a sequence, Cys-Pro ester (CPE), at the C terminus is spontaneously transformed into a peptide thioester, which can be used for the peptide ligation. In this research, we aimed to develop a method for the site-specific labeling of proteins.
At first, we made modification of the original CPE structure for the use of protein labeling, in which the ligand for protein binding was retained after thioester formation. Then, a peptide with partial sequence of parathyroid hormone (PTH) containing the modified CPE structure and a fluorescence group was synthesized. When the cell expressing a PTH receptor was treated with the peptide, synthesized, the fluorescence was observed at the same position with the receptor, and we were not able to confirm the covalent bonding between the fluorescence group and the receptor. On the other hand, we succeeded in the total chemical synthesis of proteins using the newly developed modified CPE structure.

Free Research Field

ペプチド合成