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bioRxiv · 10.1101/2021.12.30.474520

Substrate Specificity for Human Histidine Methyltransferase SETD3

Hintzen, J.·Ma, H.·Deng, H.·Witecka, A.·Andersen, S. B.·Drozak, J.·Guo, H.·Qian, P.·Li, H.·Mecinovic, J.
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Abstract

Histidine methyltransferase SETD3 plays an important role in human biology and diseases. Previously, we showed that SETD3 catalyzes N3-methylation of histidine 73 in {beta}-actin (Kwiatkowski et al., 2018). Here we report integrated synthetic, biocatalytic, biostructural and computational analyses on human SETD3-catalyzed methylation of {beta}-actin peptides possessing histidine and its structurally and chemically diverse mimics. Our enzyme assays supported by biostructural analyses demonstrate that SETD3 has a broader substrate scope beyond histidine, including N-nucleophiles on the aromatic and aliphatic side chains. Quantum mechanical/molecular mechanical (QM/MM) molecular dynamics and free-energy simulations provide insight into binding geometries and the free energy barrier for the enzymatic methyl transfer to histidine mimics, further supporting experimental data that histidine is the superior SETD3 substrate over its analogs. This work demonstrates that human SETD3 has a potential to catalyze efficient methylation of several histidine mimics, overall providing mechanistic, biocatalytic and functional insight into {beta}-actin histidine methylation by SETD3.

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Hintzen, J., Ma, H., Deng, H., Witecka, A., Andersen, S. B., Drozak, J., Guo, H., Qian, P., Li, H., Mecinovic, J.. 2021-12-30. Substrate Specificity for Human Histidine Methyltransferase SETD3. https://doi.org/10.1101/2021.12.30.474520

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