Histone demethylase that demethylates 'Lys-4' of histone H3, a specific tag for epigenetic transcriptional activation, thereby acting as a corepressor. Required for de novo DNA methylation of a subset of imprinted genes during oogenesis. Acts by oxidizing the substrate by FAD to generate the corresponding imine that is subsequently hydrolyzed. Demethylates both mono- and di-methylated 'Lys-4' of histone H3. Has no effect on tri-methylated 'Lys-4', mono-, di- or tri-methylated 'Lys-9', mono-, di- or tri-methylated 'Lys-27', mono-, di- or tri-methylated 'Lys-36' of histone H3, or on mono-, di- or tri-methylated 'Lys-20' of histone H4. Alone, it is unable to demethylate H3K4me on nucleosomes and requires the presence of GLYR1 to achieve such activity, they form a multifunctional enzyme complex that modifies transcribed chromatin and facilitates Pol II transcription through nucleosomes.
Background References
1. Marabelli C., Marrocco B., Pilotto S., Chittori S., Picaud S., Marchese S., Ciossani G., Forneris F., Filippakopoulos P., Mattevi A. A Tail-Based Mechanism Drives Nucleosome Demethylation by the LSD2/NPAC Multimeric Complex. Cell Rep. 27:387-399.e7 (2019)
2. Chen F., Yang H., Dong Z., Fang J., Wang P., Zhu T., Gong W., Fang R., Shi Y.G., Xu Y. Structural insight into substrate recognition by histone demethylase LSD2/KDM1b. Cell Res. 23:306-309 (2013)
Western blot analysis of LSD2 / AOF1 on different lysates with Rabbit anti-LSD2 / AOF1 antibody (HA722515) at 1/2,000 dilution.
Lane 1: HeLa cell lysate Lane 2: HepG2 cell lysate Lane 3: HCT 116 cell lysate Lane 4: K-562 cell lysate Lane 5: A549 cell lysate Lane 6: Jurkat cell lysate Lane 7: NIH/3T3 cell lysate Lane 8: RAW264.7 cell lysate Lane 9: Neuro-2a cell lysate Lane 10: PC-12 cell lysate Lane 11: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 92 kDa Observed band size: 92 kDa
Exposure time: 3 minutes; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722515) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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