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

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

doi:10.1016/j.pep.2020.105796

Cited by 6 publications

(5 citation statements)

References 34 publications

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“…Protein expression and purification. Throughout this work, the following TREM2 constructs are defined: TREM2 = a.a. 19-134; sTREM2 = 19-157; MBP-TREM2 = MBP fused to TREM2 19-134 as described in [18]; TREM2 receptor = 1-230. MBP-TREM2 N79Q was made by Q5 site directed mutagenesis.…”

Section: Methodsmentioning

confidence: 99%

“…We then used the results of the BLI studies with oAb42 familial variants to design a minimal Ab peptide that could be co-crystallized with TREM2, in order to structurally characterize this interaction. Since our binding results indicated that TREM2 interactions were critically mediated by the N-terminal portion of the amyloid beta peptide, we utilized the first 8 amino acids, Ab(1-8), in co-crystallization experiments with an MBP-TREM2 fusion protein that had previously been demonstrated to produce crystals that diffract to high resolution [18]. We were able to obtain crystals of the complex that diffracted to 1.84 Å (Table 1).…”

Section: Structure Of Trem2 In Complex With Ab1-8mentioning

confidence: 99%

“…Data collection and refinement statistics are given in Table 1. Crystals had similar unit cell and space group parameters to the previously published MBP-TREM2 structure [18], so the phase problem was solved by isostructural replacement using protein coordinates from PDB ID 6XDS. The initial solution was refined by rigid body refinement in PHENIX [25] and the model was improved in subsequent rounds of refinement conducted with individual B factors and iterative rebuilding through COOT [26].…”

Section: Mbp-trem2 N79q Crystallization Structure Determination and A...mentioning

confidence: 99%

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Structural and functional analysis of TREM2 interactions with amyloid beta reveal molecular mechanisms that mediate phagocytosis of oligomeric amyloid beta

Greven,

Osario,

Nix

et al. 2024

Preprint

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The TREM2 receptor is expressed on microglia in the brain, where it plays critical roles regulating microglia function. TREM2 engages a number of ligands involved in Alzheimer's disease, and consequent signaling triggers phagocytosis, activation, survival, and proliferation. TREM2 has emerged as a drug target for AD, however very little is known regarding the structural basis for TREM2 microglial functions. Here we investigated the engagement of oligomeric amyloid beta (oAb42) with TREM2. Using familial variants, we show that mutations in the N-terminal portion of Ab, notably residues H6 and D7, disrupt binding to TREM2. We then co-crystallized TREM2 with Ab(1-8) peptide and determined the high-resolution crystal structure. The structure revealed the peptide binds to the hydrophobic site of TREM2, closest to CDR1. Mutational and binding studies by BLI confirmed that mutations to the hydrophobic site ablate binding to oAb42. Finally, we show that these interactions are critical to triggering phagocytosis of oAb42, as oAb42 variants H6R and D7N are not phagocytosed. Altogether, these data indicate that TREM2 engages oAb42 using the hydrophobic site on TREM2, and the N-terminal portion of Ab, and that this interaction is critical to trigger signaling and phagocytosis.

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Section: Methodsmentioning

confidence: 99%

Section: Structure Of Trem2 In Complex With Ab1-8mentioning

confidence: 99%

Section: Mbp-trem2 N79q Crystallization Structure Determination and A...mentioning

confidence: 99%

See 1 more Smart Citation

Structural and functional analysis of TREM2 interactions with amyloid beta reveal molecular mechanisms that mediate phagocytosis of oligomeric amyloid beta

Greven,

Osario,

Nix

et al. 2024

Preprint

View full text Add to dashboard Cite

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“…However, whether this affected the immune system, metabolism and fertility and intervention at which stage of PD progression would be beneficial is unclear (133). Crystal packing interfaces analysis using the maltose binding protein (MBP)-TREM2 immunoglobulin (Ig) fusion construct has shown that the surface of the TREM2 Ig domain can bind small molecules, providing potential utility for the discovery of new therapies for TREM2 (134).…”

Section: Trem2 Treatmentmentioning

confidence: 99%

Targeting TREM2 for Parkinson’s Disease: Where to Go?

Zhang

2021

Front. Immunol.

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Parkinson’s disease (PD) is one of most common neurodegenerative disorders caused by a combination of environmental and genetic risk factors. Currently, numerous population genetic studies have shown that polymorphisms in myeloid cell-triggered receptor II (TREM2) are associated with a variety of neurodegenerative disorders. Recently, TREM2 has been verified to represent a promising candidate gene for PD susceptibility and progression. For example, the expression of TREM2 was apparently increased in the prefrontal cortex of PD patients. Moreover, the rare missense mutations in TREM2 (rs75932628, p.R47H) was confirmed to be a risk factor of PD. In addition, overexpression of TREM2 reduced dopaminergic neurodegeneration in the 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine mouse model of PD. Due to the complex pathogenesis of PD, there is still no effective drug treatment. Thus, TREM2 has received increasing widespread attention as a potential therapeutic target. This review focused on the variation of TREM2 in PD and roles of TREM2 in PD pathogenesis, such as excessive-immune inflammatory response, α-Synuclein aggregation and oxidative stress, to further provide evidence for new immune-related biomarkers and therapies for PD.

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“…There are currently eight structures of the TREM2 IgV domain publicly available (PDB IDs: 5ELI, 5UD7, 5UD8, 6B8O, 6Y6C, 6YMQ, 6YYE, and 6XDS (Byrne et al 2021;Kober et al 2016;Sudom et al 2018;Szykowska et al 2021)), as well as structures of the TM α-helix (PDB IDs: 6Z0G, 6Z0H, and 6Z0I (Steiner et al 2020)). The unliganded wild-type IgV domain (PDB IDs: 5ELI and 5UD7) is structurally similar (RMSD < 1 Å over all Cα atoms) to that observed in scFv antibody (PDB IDs: 6Y6C, 6YMQ, and 6YYE) and phosphatidylserine (PDB ID: 6B8O) complexes.…”

Section: Trem2mentioning

confidence: 99%

Structural biology of cell surface receptors implicated in Alzheimer’s disease

et al. 2021

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Alzheimer's disease is a common and devastating age-related disease with no effective disease-modifying treatments. Human genetics has implicated a wide range of cell surface receptors as playing a role in the disease, many of which are involved in the production or clearance of neurotoxins in the brain. Amyloid precursor protein, a membrane-bound signaling molecule, is at the very heart of the disease: hereditary mutations in its gene are associated with a greatly increased risk of getting the disease. A proteolytic breakdown product of amyloid precursor protein, the neurotoxic Aβ peptide, has been the target for many drug discovery efforts. Antibodies have been designed to target Aβ production with some success, although they have not proved efficacious in clinical trials with regards to cognitive benefits to date. Many of the recently identified genes associated with late-onset Alzheimer's disease risk are integral to the innate immune system. Some of these genes code for microglial proteins, such as the strongest genetic risk factor for the disease, namely APOE, and the cell surface receptors CD33 and TREM2 which are involved in clearance of the Aβ peptide from the brain. In this review, we show how structural biology has provided key insights into the normal functioning of these cell surface receptors and provided a framework for developing novel treatments to combat Alzheimer's disease.

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

Cited by 6 publications

References 34 publications

Structural and functional analysis of TREM2 interactions with amyloid beta reveal molecular mechanisms that mediate phagocytosis of oligomeric amyloid beta

Structural and functional analysis of TREM2 interactions with amyloid beta reveal molecular mechanisms that mediate phagocytosis of oligomeric amyloid beta

Targeting TREM2 for Parkinson’s Disease: Where to Go?

Structural biology of cell surface receptors implicated in Alzheimer’s disease

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