9GTV
Crystal structure of RamR with Tyr59 replaced with para-boronophenylalanine (boronate form)
This is a non-PDB format compatible entry.
Summary for 9GTV
| Entry DOI | 10.2210/pdb9gtv/pdb |
| Descriptor | Transcriptional regulator RamR (1 entity in total) |
| Functional Keywords | artificial enzyme, tetr family, boron designer enzyme, noncanonical amino acid, para-boronophenylalanine, boron acid catalysis, dna binding protein |
| Biological source | Salmonella enterica subsp. enterica serovar Typhimurium |
| Total number of polymer chains | 4 |
| Total formula weight | 92440.26 |
| Authors | Longwitz, L.,Brouwer, B.,Thunnissen, A.M.W.H.,Roelfes, G. (deposition date: 2024-09-18, release date: 2024-12-25, Last modification date: 2025-01-08) |
| Primary citation | Longwitz, L.,Kamer, M.D.,Brouwer, B.,Thunnissen, A.W.H.,Roelfes, G. Boron Designer Enzyme with a Hybrid Catalytic Dyad. Acs Catalysis, 14:18469-18476, 2024 Cited by PubMed Abstract: Genetically encoded noncanonical amino acids can introduce new-to-nature activation modes into enzymes. While these amino acids can act as catalysts on their own due to their inherent chemical properties, interactions with adjacent residues in an enzyme, such as those present in natural catalytic dyads or triads, unlock a higher potential for designer enzymes. We incorporated a boron-containing amino acid into the protein scaffold RamR to create an active enzyme for the kinetic resolution of α-hydroxythioesters. We found that a closely positioned lysine residue is crucial for the catalytic activity of the designer enzyme by forming a hybrid catalytic dyad with the boronic acid residue. The enzyme is capable of resolving differently substituted α-hydroxythioesters with good selectivities. High-resolution mass spectrometry, B NMR spectroscopy, and crystal structure analysis of the designer enzyme gave insight into the three steps of the mechanism (substrate binding, hydroxide transfer, product release). Mutations of a residue around the catalytic dyad led to a variant of the enzyme with 2-fold improvement of catalytic activity and selectivity. PubMed: 39722884DOI: 10.1021/acscatal.4c06052 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.78 Å) |
Structure validation
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