Combining MicroED and native mass spectrometry for structural discovery of enzyme-biosynthetic inhibitor complexes.

ABSTRACT: With the goal of accelerating the discovery of small molecule-protein complexes, we leverage fast, low-dose, event based electron counting microcrystal electron diffraction (MicroED) data collection and native mass spectrometry. This approach resolves structures of the epoxide-based cysteine protease inhibitor, and natural product, E-64, and its biosynthetic analogs bound to the model cysteine protease, papain. The combined structural power of MicroED and the analytical capabilities of native mass spectrometry (ED-MS) allows assignment of papain structures bound to E-64-like ligands with data obtained from crystal slurries soaked with mixtures of known inhibitors, and crude biosynthetic reactions. ED-MS further discriminates the highest-affinity ligand soaked into microcrystals from a broa

SUBMITTER: Vlahakis NW 

PROVIDER: S-EPMC11888187 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature