Crystallization (Comment) | Organism |
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mutants A76Y, A76Y/S77F, F132A/L128A, F132A/L128A/I123A, and F132A/L128A/I123A/D62A to 3.1, 2.7, 3.3, 3.35 and 3.4 A resolution, respectively. Like wildtype OPPs, all mutant structures contain 12 alpha-helices, nine of them surrounding a large central cavity and an elongated tunnel-shaped active site cavity surrounded by four alpha-helices In the crystal structure of the A76Y/S77F mutant, F77 is pushed away by Y76, thereby creating more space between those two large amino acids to accommodate the C20 product. A large F132 residue at the bottom of the tunnel-shaped active site serves as the floor and determines the final product chain length. The substitution of F132 with a small Ala, thereby removing the blockade, leads to the synthesis of a C50 product larger than that produced by the wild-type enzyme | Thermotoga maritima |
Protein Variants | Comment | Organism |
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A76Y | mutant produces only C20 cores instead of the C40 core of octaprenyl diphosphate | Thermotoga maritima |
A76Y/S77F | mutant produces only C20 cores instead of the C40 core of octaprenyl diphosphate. The A76Y/S77F mutant synthesizes a larger amount of C20 than the A76Y mutant | Thermotoga maritima |
F132A | steady-state activity 0.0051 per s. Product chain length C45 to C60 | Thermotoga maritima |
F132A/L128A | steady-state activity 0.0008 per s. Product chain length C55, C60 | Thermotoga maritima |
F132A/L128A/I123A | steady-state activity 0.00066 per s. Produuct chain length C55 to C75 | Thermotoga maritima |
F132A/L128A/I123A/D62A | products reach C95, beyond the largest chain length generated by all known trans-prenyltransferases. Steady-state activity 0.00061 per s | Thermotoga maritima |
Organism | UniProt | Comment | Textmining |
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Thermotoga maritima | Q9X1M1 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate | - |
Thermotoga maritima | 5 diphosphate + all-trans-octaprenyl diphosphate | - |
? |