One-Pot Synthesis of a Bis-Thio-Acetone Linked Ubiquitinated Histones Using 1,3-Dibromoacetone
Guo‐Chao Chu, Rui Zhao, Xiangwei Wu, Jing Shi, Yi‐Ming Li
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.
Topics & Concepts
ChemistryUbiquitinHistoneLinkerHistone octamerNucleosomeThio-Histone H2AReagentBiochemistryStereochemistryDNAOrganic chemistryComputer scienceGeneOperating systemClick Chemistry and ApplicationsProtein Degradation and InhibitorsChemical Synthesis and Analysis