Protein Folding Analysis:Protein Encapsulation within Heavy-Atom Containing Structures

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K22252
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 37:Biomolecular chemistry and related fields
• Research Institution: Kyoto University
• Principal Investigator: FUJITA DAISHI 京都大学, 高等研究院, 准教授 (20713564)
• Project Period (FY): 2019-06-28 – 2021-03-31
• Keywords: タンパク質

Outline of Final Research Achievements

The aim of this study was to stabilize and analyze the structure of a protein encapsulated within a synthetic chemical cage, i.e., a spatial modification approach.Using a Cutinase-like enzyme as a model protein, we found that the stabilization effect was remarkable against heat, denaturing agents, and organic solvents. In particular, the effect for organic solvents was more than 1000-fold. When the organic solvent ratio was increased to 90% and left the sample at room temperature for a week, the caged proteins were denatured, but no aggregation was observed. Then the refolding of the structure occurred when the solvent conditions were restored. In summary, we succeeded in stabilizing the intermediate state of folding as hypothesized.

Free Research Field

化学

Academic Significance and Societal Importance of the Research Achievements

タンパク質のフォールディング機構は、未だにわかっていないことが多い。研究の困難さの一つは、フォールディング途中の構造を、観測のために留め置くことができないためである。多くの場合、フォールディングの中途状態は疎水面が露出しているなど不安定なため、そのままでは凝集してしまう。自然界では、シャペロニンタンパク質と呼ばれる、正しいフォールディングを助ける役割のタンパク質が存在する。今回の研究は、合成した化学分子にて、シャペロニンタンパク質に相当する孤立空間を作成、狙い通りに中間状態を安定化することに成功した。