The graphene-based affinity cryo-EM grid for the endogenous protein structure determination.

ABSTRACT: Following recent advancements in cryo-electron microscopy (cryo-EM) instrumentation and software algorithms, the next bottleneck in achieving high-resolution cryo-EM structures arises from sample preparation. To overcome this, we developed a graphene-based affinity cryo-EM grid, the Graffendor (GFD) grid, to target low-abundance endogenous protein complexes. To maintain grid quality and consistency within a single batch of 36 grids, we established a one-step crosslinking batch-production method using genetically modified ALFA nanobody as affinity probe (GFD-A grid). Using low concentrations of β-galactosidase-2xALFA, we demonstrated the GFD-A grid's efficiency in capturing tagged proteins and resolving its cryo-EM structure at 2.71 Å. To test its application for endogenous proteins, we eng

SUBMITTER: An S 

PROVIDER: S-EPMC11888290 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature