MyD88 TIR Domain Higher-order Assembly Interactions Revealed by Microcrystal Electron Diffraction and Serial Femtosecond Crystallography

Overview

Journal Nat Commun

Specialty Biology

Date 2021 May 11

PMID 33972532

Citations 44

Authors

Max T B Clabbers

Susannah Holmes

Timothy W Muusse

Parimala R Vajjhala

Sara J Thygesen

Alpeshkumar K Malde

Dominic J B Hunter

Tristan I Croll

Leonie Flueckiger

Jeffrey D Nanson

Md Habibur Rahaman

Andrew Aquila

Mark S Hunter

Mengning Liang

Chun Hong Yoon

Jingjing Zhao

Nadia A Zatsepin

Brian Abbey

Emma Sierecki

Yann Gambin

Katryn J Stacey

Connie Darmanin

Bostjan Kobe

Hongyi Xu

Thomas Ve

Affiliations

Soon will be listed here.

Abstract

MyD88 and MAL are Toll-like receptor (TLR) adaptors that signal to induce pro-inflammatory cytokine production. We previously observed that the TIR domain of MAL (MAL) forms filaments in vitro and induces formation of crystalline higher-order assemblies of the MyD88 TIR domain (MyD88). These crystals are too small for conventional X-ray crystallography, but are ideally suited to structure determination by microcrystal electron diffraction (MicroED) and serial femtosecond crystallography (SFX). Here, we present MicroED and SFX structures of the MyD88 assembly, which reveal a two-stranded higher-order assembly arrangement of TIR domains analogous to that seen previously for MAL. We demonstrate via mutagenesis that the MyD88 assembly interfaces are critical for TLR4 signaling in vivo, and we show that MAL promotes unidirectional assembly of MyD88. Collectively, our studies provide structural and mechanistic insight into TLR signal transduction and allow a direct comparison of the MicroED and SFX techniques.

Citing Articles

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Comprehensive microcrystal electron diffraction sample preparation for cryo-EM.

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

Li Y, Pacoste L, Gu W, Thygesen S, Stacey K, Ve T Acta Crystallogr D Struct Biol. 2024; 80(Pt 9):699-712.

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