STAR Protocols (Jun 2021)

Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination

  • Bing-Rui Zhou,
  • Yawen Bai

Journal volume & issue
Vol. 2, no. 2
p. 100396

Abstract

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Summary: The chromatosome, a nucleosome bound to a histone H1, is the structural unit of metazoan chromatin. Determination of the high-resolution structure of the chromatosome is challenging due to the dynamic nature of H1 binding. Here, we present a protocol for purifying an optimized single-chain antibody variable fragment (scFv) that can be used to stabilize the chromatosome for single-particle cryo-EM studies. This protocol facilitates high-resolution cryo-EM structure determination of nucleosomes with a natural DNA sequence, chromatosomes, and other protein nucleosome complexes.For complete details on the use and execution of this protocol, please refer to Zhou et al. (2021).

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