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Structural Biology 2018
Volume: 4
Biochemistry & Molecular Biology Journal
Page 47
March 15-16 2018
Barcelona, Spain
10th Edition of International Conference on
Structural Biology
ε-trimethyllysine hydroxylase (TMLH) is a non-heme Fe(II)
and 2-oxoglutarate (2OG) dependent dioxygenase located
in the submitochondrial matrix. This enzyme is crucial for
the stereospecific oxidation of ε-trimethyllysine (TML) to
β-hydroxytrimethyllysine (HTML), the first step in the biosynthesis
of L-carnitine. It is proposed that the regulation of enzymatic
activity of TMLH may have more potent cardioprotective effect
than meldonium (clinically used anti-ischemia drug) that is an
inhibitor of γ-butyrobetaine hydroxylase (GBBH), the final step of
the L-carnitine production. Due to failure of the crystallographic
methods there is still lack of information about the structure of the
TMLH and especially about its active site. In this work, we applied
in silico and in vitro methods to design the possible active site of
TMLH. The structure of the TMLH was modeled using homology
modeling approach based on the closest homolog, GBBH (used as
template). However, the overall similarity between both enzymes
was slightly below 30%. Thus, various modeling softwares
were tested, and the resulting structures were optimized during
molecular dynamics simulations. This approach gave the insights
into possible enzyme fold. Next, the NMR protein-ligand binding
experiments (T1ρ, waterLOGSY and ST1D) and the enzymatic
assay (reaction monitored by 1D 1H-NMR) revealed some crucial
structure-activity relationships (SAR) that in combination with
molecular docking and previous in silico data allowed to construct
estimated active site of TMLH.
Biography
Zelencova D is a PhD student at the Riga Technical University, Latvia. She is
the Research Assistant at the Latvian Institute of Organic Synthesis: NMR
group.
Mapping the active site of epsilon-trimethyllysine hydroxylase
Zelencova D and Liepinsh E
Latvian Institute of Organic Synthesis, Latvia
Zelencova D et al., Biochem Mol biol J, Volume 4
DOI: 10.21767/2471-8084-C1-009