Description
Short Description: Recombinant Mononucleosomes H3K4me2 (EPL) consist of a 167 bp of 601 DNA and two molecules each of histones H2A that includes amino acids 1-130 (end)(accession number NP_003503.1), H2B that includes amino acids 1-126 (end)(accession number NP_003509.1), H3.2 that includes amino acids 1-136 (end)(accession number NP_066403.2) with dimethylation at lysine 4, and H4 that includes amino acids 1-103 (end)(accession number NP_003539.1). All of these histones were expressed in E Coli. The molecular weight of histone octamer is ~108 kDa. H3K4me2 (Histone H3 dimethyl Lys4) protein is generated using expressed protein ligation (EPL) technology. Truncated human Histone H3.2 is produced in E Coli and purified using FPLC. The purified protein is subsequently ligated to an N-terminal histone tail peptide containing dimethyl lysine 4 via a native peptide bond. Nucleosomes comprise the smallest subunit of chromatin; they consist of 147 bp of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for use in in vitro studies. More importantly, some histone methyltransferases are significantly more active, as well as specific, when using nucleosomal substrates in HMT assays, such as DOT1L and NSD family enzymes. Nucleosomes are also widely used in histone methyltransferase screening assays to identify small molecular inhibitors for drug discovery