AI Article Synopsis
- Histone ubiquitination is important for modifying nucleosome structures and functions.
- Researchers developed a method to create ubiquitinated histone analogues using a cross-linking reagent called 1,3-dibromoacetone (DBA).
- The method involves an efficient installation of DBA on a specific mutant of ubiquitin, followed by attaching it to histones, requiring minimal purification steps to yield significant amounts of the product.
Article Abstract
Histone ubiquitination affects the structure and function of nucleosomes. Here, we reported a one-pot synthesis of ubiquitinated histone analogues using 1,3-dibromoacetone (DBA) as the cross-linking reagent. The key step is that under the acidic borate buffer, the DBA linker can be efficiently installed to the Cys of the recombinant Ub mutant, followed by the coupling between the Ub-DBA with histones at physiological pH. The process requires a single HPLC step or orthogonal affinity tag purification to obtain ubiquitinated histone at about 24-38 mg/L expression.
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| http://dx.doi.org/10.1021/acs.joc.0c01851 | DOI Listing |
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