Investigating the active site of human trimethyllysine hydroxylase

Yali Wang, Y Vijayendar Reddy, Abbas H K Al Temimi, Hanka Venselaar, Frank H T Nelissen, Danny C Lenstra, Jasmin Mecinović

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

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.

OriginalsprogEngelsk
TidsskriftThe Biochemical journal
Vol/bind476
Udgave nummer7
Sider (fra-til)1109-1119
ISSN0264-6021
DOI
StatusUdgivet - 10. apr. 2019

Fingeraftryk

Catalytic Domain
Carnitine
Biosynthesis
Oxygenases
Hydroxylation
Mutagenesis
Amino Acid Substitution
Site-Directed Mutagenesis
trimethyllysine
trimethyl-lysine hydroxylase
Substitution reactions
Amino Acids
Substrates
alpha-ketoglutaric acid

Bibliografisk note

©2019 The Author(s).

Citer dette

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, 476(7), 1109-1119. https://doi.org/10.1042/BCJ20180857
Wang, Yali ; Reddy, Y Vijayendar ; Al Temimi, Abbas H K ; Venselaar, Hanka ; Nelissen, Frank H T ; Lenstra, Danny C ; Mecinović, Jasmin. / Investigating the active site of human trimethyllysine hydroxylase. I: The Biochemical journal. 2019 ; Bind 476, Nr. 7. s. 1109-1119.
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abstract = "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.",
author = "Yali Wang and Reddy, {Y Vijayendar} and {Al Temimi}, {Abbas H K} and Hanka Venselaar and Nelissen, {Frank H T} and Lenstra, {Danny C} and Jasmin Mecinović",
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Wang, Y, Reddy, YV, Al Temimi, AHK, Venselaar, H, Nelissen, FHT, Lenstra, DC & Mecinović, J 2019, 'Investigating the active site of human trimethyllysine hydroxylase', The Biochemical journal, bind 476, nr. 7, s. 1109-1119. https://doi.org/10.1042/BCJ20180857

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 tidsskriftTidsskriftartikelForskningpeer 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 -

Wang Y, Reddy YV, Al Temimi AHK, Venselaar H, Nelissen FHT, Lenstra DC et al. Investigating the active site of human trimethyllysine hydroxylase. The Biochemical journal. 2019 apr 10;476(7):1109-1119. https://doi.org/10.1042/BCJ20180857