7NEB
Crystal structure of branched-chain amino acid aminotransferase from Thermobaculum terrenum (M4 mutant)
Summary for 7NEB
| Entry DOI | 10.2210/pdb7neb/pdb |
| Related | 7NEA |
| Descriptor | Branched-chain-amino-acid aminotransferase, PYRIDOXAL-5'-PHOSPHATE, SODIUM ION, ... (4 entities in total) |
| Functional Keywords | transaminase, aminotransferase, bcat, iv-fold type, plp-dependent enzymes, branched-chain amino acid aminotransferases, mutant, transferase |
| Biological source | Thermobaculum terrenum (strain ATCC BAA-798 / YNP1) |
| Total number of polymer chains | 1 |
| Total formula weight | 36723.43 |
| Authors | Boyko, K.M.,Petrova, T.,Nikolaeva, A.Y.,Zeifman, Y.S.,Rakitina, T.V.,Suplatov, D.A.,Popov, V.O.,Bezsudnova, E.Y. (deposition date: 2021-02-03, release date: 2021-07-07, Last modification date: 2024-01-31) |
| Primary citation | Bezsudnova, E.Y.,Nikolaeva, A.Y.,Bakunova, A.K.,Rakitina, T.V.,Suplatov, D.A.,Popov, V.O.,Boyko, K.M. Probing the role of the residues in the active site of the transaminase from Thermobaculum terrenum. Plos One, 16:e0255098-e0255098, 2021 Cited by PubMed Abstract: Creating biocatalysts for (R)-selective amination effectively is highly desirable in organic synthesis. Despite noticeable progress in the engineering of (R)-amine activity in pyridoxal-5'-phosphate-dependent transaminases of fold type IV, the specialization of the activity is still an intuitive task, as there is poor understanding of sequence-structure-function relationships. In this study, we analyzed this relationship in transaminase from Thermobaculum terrenum, distinguished by expanded substrate specificity and activity in reactions with L-amino acids and (R)-(+)-1-phenylethylamine using α-ketoglutarate and pyruvate as amino acceptors. We performed site-directed mutagenesis to create a panel of the enzyme variants, which differ in the active site residues from the parent enzyme to a putative transaminase specific to (R)-primary amines. The variants were examined in the overall transamination reactions and half-reaction with (R)-(+)-1-phenylethylamine. A structural analysis of the most prominent variants revealed a spatial reorganization in the active sites, which caused changes in activity. Although the specialization to (R)-amine transaminase was not implemented, we succeeded in understanding the role of the particular active site residues in expanding substrate specificity of the enzyme. We showed that the specificity for (R)-(+)-1-phenylethylamine in transaminase from T. terrenum arises without sacrificing the specificity for L-amino acids and α-ketoglutarate and in consensus with it. PubMed: 34324538DOI: 10.1371/journal.pone.0255098 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.2 Å) |
Structure validation
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