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J Mol Biol. 2014 Jan 9;426(1):150-68. doi: 10.1016/j.jmb.2013.09.012. Epub 2013 Sep 17.

The N-terminal sequence of tyrosine hydroxylase is a conformationally versatile motif that binds 14-3-3 proteins and membranes.

Author information

1
Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5009 Bergen, Norway.
2
Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
3
Department of Molecular Biology, University of Bergen, Thormøhlensgate 55, N-5020 Bergen, Norway.
4
Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5009 Bergen, Norway. Electronic address: aurora.martinez@biomed.uib.no.

Abstract

Tyrosine hydroxylase (TH) catalyzes the rate-limiting step in the synthesis of catecholamine neurotransmitters, and a reduction in TH activity is associated with several neurological diseases. Human TH is regulated, among other mechanisms, by Ser19-phosphorylation-dependent interaction with 14-3-3 proteins. The N-terminal sequence (residues 1-43), which corresponds to an extension to the TH regulatory domain, also interacts with negatively charged membranes. By using X-ray crystallography together with molecular dynamics simulations and structural bioinformatics analysis, we have probed the conformations of the Ser19-phosphorylated N-terminal peptide [THp-(1-43)] bound to 14-3-3γ, free in solution and bound to a phospholipid bilayer, and of the unphosphorylated peptide TH-(1-43) both free and bilayer bound. As seen in the crystal structure of THp-(1-43) complexed with 14-3-3γ, the region surrounding pSer19 adopts an extended conformation in the bound state, whereas THp-(1-43) adopts a bent conformation when free in solution, with higher content of secondary structure and higher number of internal hydrogen bonds. TH-(1-43) in solution presents the highest mobility and least defined structure of all forms studied, and it shows an energetically more favorable interaction with membranes relative to THp-(1-43). Cationic residues, notably Arg15 and Arg16, which are the recognition sites of the kinases phosphorylating at Ser19, are also contributing to the interaction with the membrane. Our results reveal the structural flexibility of this region of TH, in accordance with the functional versatility and conformational adaptation to different partners. Furthermore, this structural information has potential relevance for the development of therapeutics for neurodegenerative disorders, through modulation of TH-partner interactions.

KEYWORDS:

MD; MM/PBSA; PC; POPS; RU; SPR; TH; X-ray crystallography; free energy of binding; molecular dynamics; molecular dynamics simulations; molecular mechanics Poisson–Boltzmann surface area; palmitoyl-oleoyl phosphatidylserine; phosphatidylcholine; phospholipid bilayers; phosphorylation; response units; surface plasmon resonance; tyrosine hydroxylase

PMID:
24055376
PMCID:
PMC3872242
DOI:
10.1016/j.jmb.2013.09.012
[Indexed for MEDLINE]
Free PMC Article
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