Development of binding scaffold with a protein from a hyperthermophilic archaeon

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K05827
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38030:Applied biochemistry-related
• Research Institution: Osaka University
• Principal Investigator: Koga Yuichi 大阪大学, 工学研究科, 准教授 (30379119)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 可溶性改変 / binding scaffold / 熱安定性

Outline of Final Research Achievements

A thermostable protein domain from a protease is expected to develop to artificial binding scaffold. To improve the solubility of SD1, SD1 protein was mutated by rational design using several structure prediction programs (foldx and Tango). The designed mutants were expressed in E. coli, and mutations of SD1 surface residues (Y15K, W18R, S106R) improved solubility expression. Furthermore, introduction of mutations (S26K, Q91P) in the aggregation-prone region (APR) identified by calculation improved soluble expression.

Free Research Field

生物工学

Academic Significance and Societal Importance of the Research Achievements

超好熱菌由来プロテアーゼの一部であるSD1は、抗体の基質結合ドメインと類似の構造をとっており、標的に対して特異的に結合する分子（binding scaffold)としての開発が期待されるが、タンパク質の可溶性に問題があった。本研究で可溶性の向上したSD1が得られたことから、SD1のbinding scaffoldとしての課題を解決したものである。