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. Author manuscript; available in PMC: 2018 Nov 21.
Published in final edited form as: Mol Biosyst. 2017 Nov 21;13(12):2509–2520. doi: 10.1039/c7mb00391a

Fig. 1. Principle of AlphaLISA™ screen for quantifying PRMT5 methylation activity.

Fig. 1

Biotinylated histone H4 is incubated with PRMT5 and a methyl donor, S-adenosyl-l-methionine (SAM). PRMT5 symmetrically dimethylates the third arginine (R3) on Biotin-H4 to form dimethylated Biotin-H4R3 (Biotin-H4R3me2) peptide. This product will further bind to Acceptor beads specific to the methylation site (Biotin-H4R3me2). Streptavidin-coated Donor beads will bind to the biotinylated substrate. Interaction between the Acceptor and Donor beads emits a chemiluminescent signal, which will be detected using an EnVision® reader. The methylation activity of PRMT5 is proportional to the intensity of the signal.