日本質量分析学会 第67回質量分析総合討論会

演題概要

オーラルセッション

第3日 5月17日(金) 16:25~16:45 C会場(101)

HDX-MSを活用した溶液中にて不安定な蛋白質の相互作用解析

(1東大医科研2東大院工)
o長門石曉1津本浩平1,2

It is difficult to predict interaction mechanisms of natural flexible and unstable proteins at the molecular level. Therefore, details on their molecular recognition mechanism, in particular physicochemical findings, still remain challenging. Here we report unknown protein-protein interactions based on biophysical analyses with hydrogen-deuterium exchange mass spectrometry (HDX-MS).
First topic is the elucidation of interactions for a target native denatured protein. By fragmenting the target protein, we identified the core regions contributing to the binding free energy by using HDX-MS and isothermal titration calorimetry (ITC). Furthermore, it is suggested that the target protein adopts a binding mode covering the entire partner protein. In this way, we succeeded in grasping the mode of interaction in solution of natural denatured protein using HDX-MSC. The second topic is the elucidation of the comprehensive interaction analysis of IL signaling complex, one of the members of the gp130 cytokine family. Thermodynamic parameters using surface plasmon resonance (SPR) and interaction mappings using HDX-MS clearly show the formation of IL- ILRα- gp130 complexes.
Thus, HDX-MS is considered to be one of the powerful analytical tools that provide useful knowledge of protein function and guidance in the search and design of inhibitors targeting proteins.