Microcrystal electron diffraction structure of Toll-like receptor 2 TIR-domain-nucleated MyD88 TIR-domain higher-order assembly

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 80; no. Pt 9; pp. 699 - 712
• Main Authors: Li, Y, Pacoste, L C, Gu, W, Thygesen, S J, Stacey, K J, Ve, T, Kobe, B, Xu, H, Nanson, J D
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.09.2024
• Subjects: Adaptor proteins; Assemblies; Assembly; Crystallography, X-Ray - methods; Electron diffraction; Humans; Microcrystals; Models, Molecular; MyD88 protein; Myeloid Differentiation Factor 88 - chemistry; Myeloid Differentiation Factor 88 - metabolism; Protein Domains; Protein Multimerization; Proteins; Receptors; Receptors, Interleukin-1 - chemistry; Receptors, Interleukin-1 - metabolism; TLR1 protein; TLR2 protein; Toll-Like Receptor 1 - chemistry; Toll-Like Receptor 1 - metabolism; Toll-Like Receptor 2 - chemistry; Toll-Like Receptor 2 - metabolism; Toll-Like Receptor 6 - chemistry; Toll-Like Receptor 6 - metabolism; Toll-like receptors; higher-order assemblies; MicroED; Toll-like receptors; signalosomes; MyD88; Toll/interleukin-1 receptor protein; TIR domains
• ISSN: 0907-4449; 2059-7983; 1399-0047
• DOI: 10.1107/S2059798324008210

Eukaryotic TIR (Toll/interleukin-1 receptor protein) domains signal via TIR-TIR interactions, either by self-association or by interaction with other TIR domains. In mammals, TIR domains are found in Toll-like receptors (TLRs) and cytoplasmic adaptor proteins involved in pro-inflammatory signaling. Previous work revealed that the MAL TIR domain (MAL ) nucleates the assembly of MyD88 into crystalline arrays in vitro. A microcrystal electron diffraction (MicroED) structure of the MyD88 assembly has previously been solved, revealing a two-stranded higher-order assembly of TIR domains. In this work, it is demonstrated that the TIR domain of TLR2, which is reported to signal as a heterodimer with either TLR1 or TLR6, induces the formation of crystalline higher-order assemblies of MyD88 in vitro, whereas TLR1 and TLR6 do not. Using an improved data-collection protocol, the MicroED structure of TLR2 -induced MyD88 microcrystals was determined at a higher resolution (2.85 Å) and with higher completeness (89%) compared with the previous structure of the MAL -induced MyD88 assembly. Both assemblies exhibit conformational differences in several areas that are important for signaling (for example the BB loop and CD loop) compared with their monomeric structures. These data suggest that TLR2 and MAL interact with MyD88 in an analogous manner during signaling, nucleating MyD88 assemblies unidirectionally.