Size-Selective VAILase Proteolysis Provides Dynamic Insights into Protein Structures.

Monitoring the dynamic alterations of protein structures within an aqueous solution remains enormously challenging. In this study, we describe a size-selective VAILase proteolysis (SVP)-mass spectrometry (MS) strategy to probe the protein structure changes without strict control of the proteolysis kinetics. The unique conformation selectivity of SVP depends on the uniform nano-sized entrance pores of the VAILase hexameric cage as well as the six inherent molecular rulers in the VAILase-substrate recognition and cleavage. The dynamic insights into subtle conformation alterations of both myoglobin unfolding transition and Aurora kinase A-inhibitor binding are successfully captured using the SVP strategy, which matches well with the results in the molecular dynamics simulation. Our work provides a new paradigm of size-selective native proteolysis for exploring the aqueous protein structure-function relationships.

Citation

Binwen Sun, Ji Lv, Jin Chen, Zheyi Liu, Ye Zhou, Lin Liu, Yan Jin, Fangjun Wang. Size-Selective VAILase Proteolysis Provides Dynamic Insights into Protein Structures. Analytical chemistry. 2021 Aug 03;93(30):10653-10660

Mesh Tags

Substances

PMID: 34291915

search → result