Methyltransferase Assay Services

Creative Biolabs provides methyltransferase assay services in an effort to alleviate the challenges involved in locating epigenetic medications. Our clients may build an early discovery pipeline around methyltransferase targets thanks to our platform technology and highly skilled personnel.

Protein methyltransferases (PMTs) are members of the transferase enzyme family, which also includes kinases, acetyltransferases, and glycosyltransferases. Transferring a functional group from a source (cofactor or coenzyme) to an acceptor is what unites these enzymes. S-adenosylmethionine (SAM) and the lysine or arginine side chains of protein substrates serve as the cofactor and acceptor, respectively, for PMTs. More than 60 PMTs, including more than 50 protein lysine methyltransferases (PKMTs) and 9 recognized protein arginine methyltransferases (PRMTs), are encoded by the human genome. The THW loop for SAM binding and a collection of four conserved motifs are shared by the nine human PRMTs (PRMT1–9). PRMTs target-specifically alter the ω-guanidino nitrogen of arginine using SAM as the methyl donor. For the purpose of binding SAM and catalyzing enzymes, PKMTs have a canonical SET domain consisting of 130 amino acids.

Fig.1. Reactions catalyzed by a PMT.¹

Targets of Methyltransferase Assay Services

In addition to other posttranslational alterations, PMT-mediated histone and nonhistone methylation can control the binding partners, stability, and localization of PMT substrates. These changes, either separately or in combination, have the ability to epigenetically modify downstream signals. In addition to their functions in healthy physiology, PMT dysregulation has been linked to a number of illnesses, including cancer. PMTs' carcinogenic qualities may be due to target methylation, which can inhibit or destabilize tumor suppressors. Targeting numerous PMTs, Creative Biolabs is able to provide methyltransferase assay services:

Substrates of PMTs in Methyltransferase Assay Services

• Peptides as PMT Substrates

Both the matching peptides and protein substrates can be recognized by a large number of PMTs. Peptides and their derivatives have been widely employed as in vitro substrates to characterize PMTs because they are easily produced via solid-phase peptide synthesis. Using PRMT1 as an example, its methylation activities (k_cat/K_m) on histone H4 and the N-terminal H4 1-21 peptide are comparable; however, they drop 200-fold when it comes to the N-terminal H4 1-18 peptide and the equivalent R19A peptide.

• Proteins or Protein Complexes as PMT Substrates

The type of substrates that PMTs use can have a significant impact on their target specificity. For example, when nucleosomes are available as substrates, NSD2 methylates H3K36; whereas, when histone octamers are available as substrates, NSD2 operates on H4K44. Full-length proteins or protein complexes are more pertinent in these situations as PMT in vitro substrates.

Creative Biolabs has concentrated on PMTs in disorders and epigenetics. We are actively working on creating tools to clarify and control PMT-related methylation. Our scientific staff is committed to making sure that the best possible methyltransferase assay services are provided. Please do not hesitate to contact us if you have any queries or extra requests regarding the methyltransferase assay.

Reference

1. Luo, Minkui. "Current chemical biology approaches to interrogate protein methyltransferases." ACS chemical biology 7.3 (2012): 443-463.