期刊
CHEMBIOCHEM
卷 23, 期 1, 页码 -出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.202100437
关键词
archaeal MAT; crystal structures; MjMAT; photocaging; SAM synthetase
资金
- DFG [SFB858, CRC1459, 433682494]
- European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme [772280]
- HZB
- Projekt DEAL
Methylation and demethylation of DNA, RNA, and proteins play a key role in epigenetic regulation. Engineered methionine adenosyltransferases have been utilized to modify biomolecules with non-natural groups, leading to efficient conversion of specific substrates and enabling photo-protection by visible light.
Methylation and demethylation of DNA, RNA and proteins constitutes a major regulatory mechanism in epigenetic processes. Investigations would benefit from the ability to install photo-cleavable groups at methyltransferase target sites that block interactions with reader proteins until removed by non-damaging light in the visible spectrum. Engineered methionine adenosyltransferases (MATs) have been exploited in cascade reactions with methyltransferases (MTases) to modify biomolecules with non-natural groups, including first evidence for accepting photo-cleavable groups. We show that an engineered MAT from Methanocaldococcus jannaschii (PC-MjMAT) is 308-fold more efficient at converting ortho-nitrobenzyl-(ONB)-homocysteine than the wildtype enzyme. PC-MjMAT is active over a broad range of temperatures and compatible with MTases from mesophilic organisms. We solved the crystal structures of wildtype and PC-MjMAT in complex with AdoONB and a red-shifted derivative thereof. These structures reveal that aromatic stacking interactions within the ligands are key to accommodating the photocaging groups in PC-MjMAT. The enlargement of the binding pocket eliminates steric clashes to enable AdoMet analogue binding. Importantly, PC-MjMAT exhibits remarkable activity on methionine analogues with red-shifted ONB-derivatives enabling photo-deprotection of modified DNA by visible light.
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