Table 1: Methods Used in the Discovery of Small Molecule Inhibitors of Histone Methyltransferase and Demethylase Enzymes

Method Compound Target Secondary Assays
Radiotracer Assays
Filter Binding Chaetocin [6] SU(VAR)3-9 Biochemical and cellular immunoblot, MS
Pyrazole Amide 7b [68] PRMT4 PRMT1, PRMT3 selectivity (filter binding)
[14C]-Labeling Sinefungin [69] N.D. N.D.
Antibody-Based Assays
DELFIA BIX-01294, BIX-01338 [13, 32] G9a, GLP MS, Cellular expression analysis, ChIP
ELISA AMI-1, AMI-5 [12] Hmt1p, PRMT1 Filter binding, ERα-reporter gene assay
Coupled Enzyme Assays
FDH-Coupled Myricetin, β-lapachone [45] JMJD2E MS
8-hydroxyquinolines [43] JMJD2E, 2A MS, FRAP
Mass Spectrometry (MS)
Disulfiram or selenium derivatives [70] JMJD2A Zn(II)-ejection assay
Fluorescence Polarization (FP)
Stilbamidine, allantodapsone, RM-65 [14, 71] RmtA, PRMT1 TRF, ERα -reporter gene assay, cellular IHC
Crystallography/Modeling/Virtual Screening
UNC-0321 [17] G9a, GLP SAHH-coupled assay, AlphaScreen, FP, MCE
E70 [72] GLP MS, ITC, cellular expression analysis
Tranylcypromine-Lys conjugates [73] LSD1 HRP-coupled assay, cellular immunoblot
2,4-PDCA [33] JMJD2E, 2A FDH-coupled assay, MS, crystallography
4-dimethyl-amino-benzyl-NOG [74] JMJD2A, 2C, 2D Biochemical and cellular immunoblot
N-oxalyl-D-tyrosine derivatives [75] JMJD2A MS, FDH-coupled assay, crystallography
Structural or Mechanistic Similarity
Tranylcypromine [48] LSD1 Immunoblot, ChIP, transcriptional analysis
Lys4-cyclopropyl and propargylamine H3 peptide derivatives [50] LSD1 HRP-coupled assay, MS
Bisguanidine and biguanide polyamine analogs [54] LSD1 Biochemical and cellular immunoblot, transcriptional analysis, ChIP
N-oxalylglycine (NOG) [76] JMJD2E, 2a, 2B Immunoblot, FDH-coupled assay
Succinic acid [77] JMJD2D Biochemical and cellular immunoblot

N.D., not determined.