Resumé
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 (2 S )- Nε-trimethyllysine to (2 S ,3 S )-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 (2 S )- Nε-trimethyllysine substrate provide a basic insight into the binding requirements that determine an efficient TMLH-catalyzed conversion of (2 S )- Nε-trimethyllysine to (2 S ,3 S )-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 (2 S )- Nε-trimethyllysine.
| Originalsprog | Engelsk |
|---|---|
| Tidsskrift | The Biochemical journal |
| Vol/bind | 476 |
| Udgave nummer | 7 |
| Sider (fra-til) | 1109-1119 |
| ISSN | 0264-6021 |
| DOI | |
| Status | Udgivet - 10. apr. 2019 |
Fingeraftryk
Bibliografisk note
©2019 The Author(s).Citer dette
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Investigating the active site of human trimethyllysine hydroxylase. / Wang, Yali; Reddy, Y Vijayendar; Al Temimi, Abbas H K; Venselaar, Hanka; Nelissen, Frank H T; Lenstra, Danny C; Mecinović, Jasmin.
I: The Biochemical journal, Bind 476, Nr. 7, 10.04.2019, s. 1109-1119.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review
TY - JOUR
T1 - Investigating the active site of human trimethyllysine hydroxylase
AU - Wang, Yali
AU - Reddy, Y Vijayendar
AU - Al Temimi, Abbas H K
AU - Venselaar, Hanka
AU - Nelissen, Frank H T
AU - Lenstra, Danny C
AU - Mecinović, Jasmin
N1 - ©2019 The Author(s).
PY - 2019/4/10
Y1 - 2019/4/10
N2 - 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 (2 S )- Nε-trimethyllysine to (2 S ,3 S )-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 (2 S )- Nε-trimethyllysine substrate provide a basic insight into the binding requirements that determine an efficient TMLH-catalyzed conversion of (2 S )- Nε-trimethyllysine to (2 S ,3 S )-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 (2 S )- Nε-trimethyllysine.
AB - 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 (2 S )- Nε-trimethyllysine to (2 S ,3 S )-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 (2 S )- Nε-trimethyllysine substrate provide a basic insight into the binding requirements that determine an efficient TMLH-catalyzed conversion of (2 S )- Nε-trimethyllysine to (2 S ,3 S )-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 (2 S )- Nε-trimethyllysine.
U2 - 10.1042/BCJ20180857
DO - 10.1042/BCJ20180857
M3 - Journal article
C2 - 30898847
VL - 476
SP - 1109
EP - 1119
JO - Biochemical Journal
JF - Biochemical Journal
SN - 0264-6021
IS - 7
ER -