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

Crystallization / methods Humans Maltose-Binding Proteins / chemistry Membrane Glycoproteins / chemistry Molecular Chaperones / chemistry Receptors, Immunologic / chemistry Recombinant Fusion Proteins / chemistry
Alzheimer's disease Crystallization chaperone Fusion Maltose binding protein TREM2

Journal

Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496

Informations de publication

Date de publication:
03 2021
Historique:
received: 16 06 2020
revised: 15 09 2020
accepted: 13 11 2020
pubmed: 23 11 2020
medline: 19 8 2021
entrez: 22 11 2020
Statut: ppublish

Résumé

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.

Identifiants

DOI: 10.1016/j.pep.2020.105796 PMID: 33221505
pubmed: 33221505
pii: S1046-5928(20)30387-9
doi: 10.1016/j.pep.2020.105796
pii:
doi:

Substances chimiques

Maltose-Binding Proteins 0
Membrane Glycoproteins 0
Molecular Chaperones 0
Receptors, Immunologic 0
Recombinant Fusion Proteins 0
TREM2 protein, human 0

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

105796

Informations de copyright

Copyright © 2020 Elsevier Inc. All rights reserved.

Auteurs

Noel J Byrne (NJ)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA. Electronic address: noel_byrne3@merck.com.

Amy C Lee (AC)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

James Kostas (J)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

John C Reid (JC)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

Andrea T Partridge (AT)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

Sung-Sau So (SS)

Department of Computational and Structural Sciences, Merck & Co., Inc., Kenilworth, NJ, USA.

Joseph E Cowan (JE)

Department of Discovery Sample Management, Merck & Co., Inc., Rahway, NJ, USA.

Pravien Abeywickrema (P)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

Hua Huang (H)

Evotec, Ltd., Princeton, NJ, USA.

Matthias Zebisch (M)

Evotec, Ltd., Abingdon, Oxfordshire, UK.

John J Barker (JJ)

Evotec, Ltd., Abingdon, Oxfordshire, UK.

Stephen M Soisson (SM)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

Alexei Brooun (A)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

Hua-Poo Su (HP)

Department of Computational and Structural Sciences, Merck & Co., Inc., West Point, PA, USA.

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Classifications MeSH

questionsmedicales.fr › Phénomènes chimiques › Cristallisation › Development of a robust crystallization...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Development of a robust crystallization...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Protéines de liaison au maltose › Development of a robust crystallization...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines membranaires › Glycoprotéines membranaires › Development of a robust crystallization...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Chaperons moléculaires › Development of a robust crystallization...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines membranaires › Récepteurs de surface cellulaire › Récepteurs immunologiques › Development of a robust crystallization...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines recombinantes › Protéines de fusion recombinantes › Development of a robust crystallization...