Investigating the active site of human trimethyllysine hydroxylase
| Autor: | Hanka Venselaar, Jasmin Mecinović, Frank H. T. Nelissen, Yali Wang, Abbas H. K. Al Temimi, Y. Vijayendar Reddy, Danny C. Lenstra |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Models
Molecular Oxygenase Stereochemistry gamma-Butyrobetaine Dioxygenase Synthetic Organic Chemistry 01 natural sciences Biochemistry Cofactor Mixed Function Oxygenases Substrate Specificity Hydroxylation 03 medical and health sciences chemistry.chemical_compound All institutes and research themes of the Radboud University Medical Center Carnitine Catalytic Domain Humans Amino Acid Sequence Molecular Biology 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology Sequence Homology Amino Acid 010405 organic chemistry Mutagenesis Active site Cell Biology Recombinant Proteins 0104 chemical sciences Amino acid Kinetics Enzyme chemistry Amino Acid Substitution Carnitine biosynthesis biology.protein Biocatalysis Mutagenesis Site-Directed Nanomedicine Radboud Institute for Molecular Life Sciences [Radboudumc 19] |
| Zdroj: | Biochemical Journal, 476, 1109-1119 Wang, Y, Reddy, Y V, Al Temimi, A H K, Venselaar, H, Nelissen, F H T, Lenstra, D C & Mecinović, J 2019, ' Investigating the active site of human trimethyllysine hydroxylase ', The Biochemical journal, vol. 476, no. 7, pp. 1109-1119 . https://doi.org/10.1042/BCJ20180857 Biochemical Journal, 476, pp. 1109-1119 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/BCJ20180857 |
| Popis: | The biologically important carnitine biosynthesis pathway in humans proceeds via four enzymatic steps. The first step in carnitine biosynthesis is catalyzed by trimethyllysine hydroxylase (TMLH), a non-heme Fe(II) and 2-oxoglutarate (2OG)-dependent oxygenase, which catalyzes the stereospecific hydroxylation of (2S)-Nε-trimethyllysine to (2S,3S)-3-hydroxy-Nε-trimethyllysine. Here, we report biocatalytic studies on human TMLH and its 19 variants introduced through site-directed mutagenesis. Amino acid substitutions at the sites involved in binding of the Fe(II) cofactor, 2OG cosubstrate and (2S)-Nε-trimethyllysine substrate provide a basic insight into the binding requirements that determine an efficient TMLH-catalyzed conversion of (2S)-Nε-trimethyllysine to (2S,3S)-3-hydroxy-Nε-trimethyllysine. This work demonstrates the importance of the recognition sites that contribute to the enzymatic activity of TMLH: the Fe(II)-binding H242–D244–H389 residues, R391–R398 involved in 2OG binding and several residues (D231, N334 and the aromatic cage comprised of W221, Y217 and Y234) associated with binding of (2S)-Nε-trimethyllysine. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|