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J Biomol NMR. 2019 Jun 26. doi: 10.1007/s10858-019-00257-1. [Epub ahead of print]

A unified structural model of the mammalian translocator protein (TSPO).

Author information

1
Center for Structural Biology, Vanderbilt University, Nashville, TN, 37240, USA.
2
Department of Chemistry, Vanderbilt University, Nashville, TN, 37235, USA.
3
Institute of Imaging Science, Vanderbilt University Medical Center, Nashville, TN, 37232, USA.
4
Department of Biochemistry, Vanderbilt University, Nashville, TN, 37240, USA.
5
Center for Structural Biology, Vanderbilt University, Nashville, TN, 37240, USA. jens.meiler@vanderbilt.edu.
6
Department of Chemistry, Vanderbilt University, Nashville, TN, 37235, USA. jens.meiler@vanderbilt.edu.
7
Department of Chemistry, Center for Structural Biology, Vanderbilt University, MRBIII 5144B, Nashville, TN, 37232, USA. jens.meiler@vanderbilt.edu.

Abstract

The translocator protein (TSPO), previously known as the peripheral benzodiazepine receptor (PBR), is a membrane protein located on the outer mitochondrial membrane. Experimentally-derived structures of mouse TSPO (mTSPO) and its homologs from bacterial species have been determined by NMR spectroscopy and X-ray crystallography, respectively. These structures and ligand interactions within the TSPO binding pocket display distinct differences. Here, we leverage experimental and computational studies to derive a unified structural model of mTSPO in the presence and absence of the TSPO ligand, PK11195, and study the effects of DPC detergent micelles on the TSPO structure and ligand binding. From this work, we conclude that that the lipid-mimetic system used to solubilize mTSPO for NMR studies thermodynamically destabilizes the protein, introduces structural perturbations, and alters the characteristics of ligand binding. Furthermore, we used Rosetta to construct a unified mTSPO model that reconciles deviating features of the mammalian and bacterial TSPO. These deviating features are likely a consequence of the detergent system used for structure determination of mTSPO by NMR. The unified mTSPO model agrees with available experimental NMR data, appears to be physically realistic (i.e. thermodynamically not frustrated as judged by the Rosetta energy function), and simultaneously shares the structural features observed in sequence-conserved regions of the bacterial proteins. Finally, we identified the binding site for an imaging ligand VUIIS8310 that is currently positioned for clinical translation using NMR spectroscopy and propose a computational model of the VUIIS8310-mTSPO complex.

KEYWORDS:

Homology modeling; Ligand docking; NMR spectroscopy; Protein folding; Rosetta; Translocator protein (TSPO)

PMID:
31243635
DOI:
10.1007/s10858-019-00257-1

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