Transition state for the NSD2-catalyzed methylation of histone H3 lysine 36
| Autor: | Rosalie Matico, Wangfang Hou, Thomas D. Meek, Sara H. Thrall, Patrick McDevitt, Jessica L. Schneck, Johnson Neil W, Myles B. Poulin, Michael J. Huddleston, Vern L. Schramm |
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| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular 0301 basic medicine Reaction mechanism Stereochemistry Methylation Catalysis Histones 03 medical and health sciences Histone H3 Nucleophile Kinetic isotope effect Humans Nucleosome Multidisciplinary 030102 biochemistry & molecular biology Chemistry Lysine digestive oral and skin physiology Leaving group Histone-Lysine N-Methyltransferase Biological Sciences Repressor Proteins 030104 developmental biology Biochemistry SN2 reaction HeLa Cells |
| Zdroj: | Proceedings of the National Academy of Sciences. 113:1197-1201 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1521036113 |
| Popis: | Nuclear receptor SET domain containing protein 2 (NSD2) catalyzes the methylation of histone H3 lysine 36 (H3K36). It is a determinant in Wolf-Hirschhorn syndrome and is overexpressed in human multiple myeloma. Despite the relevance of NSD2 to cancer, there are no potent, selective inhibitors of this enzyme reported. Here, a combination of kinetic isotope effect measurements and quantum chemical modeling was used to provide subangstrom details of the transition state structure for NSD2 enzymatic activity. Kinetic isotope effects were measured for the methylation of isolated HeLa cell nucleosomes by NSD2. NSD2 preferentially catalyzes the dimethylation of H3K36 along with a reduced preference for H3K36 monomethylation. Primary Me-(14)C and (36)S and secondary Me-(3)H3, Me-(2)H3, 5'-(14)C, and 5'-(3)H2 kinetic isotope effects were measured for the methylation of H3K36 using specifically labeled S-adenosyl-l-methionine. The intrinsic kinetic isotope effects were used as boundary constraints for quantum mechanical calculations for the NSD2 transition state. The experimental and calculated kinetic isotope effects are consistent with an SN2 chemical mechanism with methyl transfer as the first irreversible chemical step in the reaction mechanism. The transition state is a late, asymmetric nucleophilic displacement with bond separation from the leaving group at (2.53 Å) and bond making to the attacking nucleophile (2.10 Å) advanced at the transition state. The transition state structure can be represented in a molecular electrostatic potential map to guide the design of inhibitors that mimic the transition state geometry and charge. |
| Databáze: | OpenAIRE |
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