Development of a robust crystallization platform for immune receptor TREM2 using a crystallization chaperone strategy

Noel J Byrne; Amy C Lee; James Kostas; John C Reid; Andrea T Partridge; Sung-Sau So; Joseph E Cowan; Pravien Abeywickrema; Hua Huang; Matthias Zebisch; John J Barker; Stephen M Soisson; Alexei Brooun; Hua-Poo Su

doi:10.1016/j.pep.2020.105796

Development of a robust crystallization platform for immune receptor TREM2 using a crystallization chaperone strategy

Protein Expr Purif. 2021 Mar:179:105796. doi: 10.1016/j.pep.2020.105796. Epub 2020 Nov 20.

Authors

Affiliations

¹ Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA. Electronic address: noel_byrne3@merck.com.
² Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.
³ Department of Computational and Structural Sciences, Merck & Co., Inc., Kenilworth, NJ, USA.
⁴ Department of Discovery Sample Management, Merck & Co., Inc., Rahway, NJ, USA.
⁵ Evotec, Ltd., Princeton, NJ, USA.
⁶ Evotec, Ltd., Abingdon, Oxfordshire, UK.

PMID: 33221505
DOI: 10.1016/j.pep.2020.105796

Abstract

TREM2 has been identified by genomic analysis as a potential and novel target for the treatment of Alzheimer's disease. To enable structure-based screening of potential small molecule therapeutics, we sought to develop a robust crystallization platform for the TREM2 Ig-like domain. A systematic set of constructs containing the structural chaperone, maltose binding protein (MBP), fused to the Ig domain of TREM2, were evaluated in parallel expression and purification, followed by crystallization studies. Using protein crystallization and high-resolution diffraction as a readout, a MBP-TREM2 Ig fusion construct was identified that generates reproducible protein crystals diffracting at 2.0 Å, which makes it suitable for soaking of potential ligands. Importantly, analysis of crystal packing interfaces indicates that most of the surface of the TREM2 Ig domain is available for small molecule binding. A proof of concept co-crystallization study with a small library of fragments validated potential utility of this system for the discovery of new TREM2 therapeutics.

Keywords: Alzheimer's disease; Crystallization chaperone; Fusion; Maltose binding protein; TREM2.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Crystallization / methods*
Humans
Maltose-Binding Proteins / chemistry
Maltose-Binding Proteins / genetics
Maltose-Binding Proteins / metabolism
Membrane Glycoproteins* / chemistry
Membrane Glycoproteins* / genetics
Membrane Glycoproteins* / metabolism
Molecular Chaperones* / chemistry
Molecular Chaperones* / metabolism
Receptors, Immunologic* / chemistry
Receptors, Immunologic* / genetics
Receptors, Immunologic* / metabolism
Recombinant Fusion Proteins* / chemistry
Recombinant Fusion Proteins* / genetics
Recombinant Fusion Proteins* / metabolism

Substances

Maltose-Binding Proteins
Membrane Glycoproteins
Molecular Chaperones
Receptors, Immunologic
Recombinant Fusion Proteins
TREM2 protein, human