6P9V
Crystal Structure of hMAT Mutant K289L
Summary for 6P9V
| Entry DOI | 10.2210/pdb6p9v/pdb |
| Descriptor | S-adenosylmethionine synthase isoform type-2, ADENOSINE, POTASSIUM ION, ... (6 entities in total) |
| Functional Keywords | adenosyl transferase, adomet synthase, natural product enzyme, transferase |
| Biological source | Homo sapiens (Human) |
| Total number of polymer chains | 1 |
| Total formula weight | 46382.55 |
| Authors | Miller, M.D.,Xu, W.,Huber, T.D.,Clinger, J.A.,Liu, Y.,Thorson, J.S.,Phillips Jr., G.N. (deposition date: 2019-06-10, release date: 2020-04-22, Last modification date: 2023-10-11) |
| Primary citation | Huber, T.D.,Clinger, J.A.,Liu, Y.,Xu, W.,Miller, M.D.,Phillips Jr., G.N.,Thorson, J.S. Methionine Adenosyltransferase Engineering to Enable Bioorthogonal Platforms for AdoMet-Utilizing Enzymes. Acs Chem.Biol., 15:695-705, 2020 Cited by PubMed Abstract: The structural conservation among methyltransferases (MTs) and MT functional redundancy is a major challenge to the cellular study of individual MTs. As a first step toward the development of an alternative biorthogonal platform for MTs and other AdoMet-utilizing enzymes, we describe the evaluation of 38 human methionine adenosyltransferase II-α (hMAT2A) mutants in combination with 14 non-native methionine analogues to identify suitable bioorthogonal mutant/analogue pairings. Enabled by the development and implementation of a hMAT2A high-throughput (HT) assay, this study revealed hMAT2A K289L to afford a 160-fold inversion of the hMAT2A selectivity index for a non-native methionine analogue over the native substrate l-Met. Structure elucidation of K289L revealed the mutant to be folded normally with minor observed repacking within the modified substrate pocket. This study highlights the first example of exchanging l-Met terminal carboxylate/amine recognition elements within the hMAT2A active-site to enable non-native bioorthgonal substrate utilization. Additionally, several hMAT2A mutants and l-Met substrate analogues produced AdoMet analogue products with increased stability. As many AdoMet-producing (e.g., hMAT2A) and AdoMet-utlizing (e.g., MTs) enzymes adopt similar active-site strategies for substrate recognition, the proof of concept first generation hMAT2A engineering highlighted herein is expected to translate to a range of AdoMet-utilizing target enzymes. PubMed: 32091873DOI: 10.1021/acschembio.9b00943 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.051 Å) |
Structure validation
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