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- EMDB-11983: Cryo-EM of Aedes Aegypti Toll5A trimer bound to Spz1C -

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Basic information

Entry
Database: EMDB / ID: EMD-11983
TitleCryo-EM of Aedes Aegypti Toll5A trimer bound to Spz1C
Map dataMap
Sample
  • Complex: Toll5A-Spz1C complex
    • Complex: Toll-like receptor
      • Protein or peptide: Toll-like receptor
    • Complex: AAEL013433-PA
      • Protein or peptide: AAEL013433-PA
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
KeywordsMosquito vector biology / Toll receptor / LRR / Cystine knot domain / IMMUNE SYSTEM
Function / homology
Function and homology information


Toll binding / central nervous system formation / toll-like receptor signaling pathway / growth factor activity / transmembrane signaling receptor activity / inflammatory response / immune response / innate immune response / extracellular space / membrane
Similarity search - Function
Spaetzle / : / Spaetzle / Toll-like receptor / TIR domain / Leucine-rich repeats, bacterial type / Toll - interleukin 1 - resistance / Cystine-knot cytokine / TIR domain profile. / Leucine-rich repeat, SDS22-like subfamily ...Spaetzle / : / Spaetzle / Toll-like receptor / TIR domain / Leucine-rich repeats, bacterial type / Toll - interleukin 1 - resistance / Cystine-knot cytokine / TIR domain profile. / Leucine-rich repeat, SDS22-like subfamily / Toll/interleukin-1 receptor homology (TIR) domain / Toll/interleukin-1 receptor homology (TIR) domain superfamily / Leucine rich repeat / Leucine-rich repeat, typical subtype / Leucine-rich repeats, typical (most populated) subfamily / Leucine-rich repeat profile. / Leucine-rich repeat / Leucine-rich repeat domain superfamily
Similarity search - Domain/homology
TIR domain-containing protein / AAEL013433-PA
Similarity search - Component
Biological speciesAedes aegypti (yellow fever mosquito)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.74 Å
AuthorsGangloff M / Hardwick SW / Chirgadze DY
Funding support United Kingdom, 1 items
OrganizationGrant numberCountry
Medical Research Council (MRC, United Kingdom)MR/P02260X/1 United Kingdom
CitationJournal: Nat Commun / Year: 2022
Title: Structure and dynamics of Toll immunoreceptor activation in the mosquito Aedes aegypti.
Authors: Yoann Saucereau / Thomas H Wilson / Matthew C K Tang / Martin C Moncrieffe / Steven W Hardwick / Dimitri Y Chirgadze / Sandro G Soares / Maria Jose Marcaida / Nicholas J Gay / Monique Gangloff /
Abstract: Aedes aegypti has evolved to become an efficient vector for arboviruses but the mechanisms of host-pathogen tolerance are unknown. Immunoreceptor Toll and its ligand Spaetzle have undergone ...Aedes aegypti has evolved to become an efficient vector for arboviruses but the mechanisms of host-pathogen tolerance are unknown. Immunoreceptor Toll and its ligand Spaetzle have undergone duplication which may allow neofunctionalization and adaptation. Here we present cryo-EM structures and biophysical characterisation of low affinity Toll5A complexes that display transient but specific interactions with Spaetzle1C, forming asymmetric complexes, with only one ligand clearly resolved. Loop structures of Spaetzle1C and Toll5A intercalate, temporarily bridging the receptor C-termini to promote signalling. By contrast unbound receptors form head-to-head homodimers that keep the juxtamembrane regions far apart in an inactive conformation. Interestingly the transcriptional signature of Spaetzle1C differs from other Spaetzle cytokines and controls genes involved in innate immunity, metabolism and tissue regeneration. Taken together our results explain how upregulation of Spaetzle1C in the midgut and Toll5A in the salivary gland shape the concomitant immune response.
History
DepositionNov 24, 2020-
Header (metadata) releaseJul 13, 2022-
Map releaseJul 13, 2022-
UpdateOct 23, 2024-
Current statusOct 23, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_11983.map.gz / Format: CCP4 / Size: 209.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationMap
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.83 Å/pix.
x 380 pix.
= 315.4 Å
0.83 Å/pix.
x 380 pix.
= 315.4 Å
0.83 Å/pix.
x 380 pix.
= 315.4 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.83 Å
Density
Contour LevelBy AUTHOR: 0.08
Minimum - Maximum-0.11554735 - 0.36830458
Average (Standard dev.)0.0009553882 (±0.013300346)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions380380380
Spacing380380380
CellA=B=C: 315.4 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Half map

Fileemd_11983_half_map_1.map
AnnotationHalf map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: Half map

Fileemd_11983_half_map_2.map
AnnotationHalf map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Toll5A-Spz1C complex

EntireName: Toll5A-Spz1C complex
Components
  • Complex: Toll5A-Spz1C complex
    • Complex: Toll-like receptor
      • Protein or peptide: Toll-like receptor
    • Complex: AAEL013433-PA
      • Protein or peptide: AAEL013433-PA
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: Toll5A-Spz1C complex

SupramoleculeName: Toll5A-Spz1C complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 / Details: Holodimer complex of Toll5A bound to Spz1C
Molecular weightTheoretical: 230 kDa/nm

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Supramolecule #2: Toll-like receptor

SupramoleculeName: Toll-like receptor / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1
Source (natural)Organism: Aedes aegypti (yellow fever mosquito)

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Supramolecule #3: AAEL013433-PA

SupramoleculeName: AAEL013433-PA / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2
Source (natural)Organism: Aedes aegypti (yellow fever mosquito)

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Macromolecule #1: Toll-like receptor

MacromoleculeName: Toll-like receptor / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Aedes aegypti (yellow fever mosquito)
Molecular weightTheoretical: 87.675773 KDa
Recombinant expressionOrganism: Drosophila melanogaster (fruit fly)
SequenceString: TSTKRFTCPE ESEASNCSCE EFPSKTHFYC PDFNPTLYVD VEDRMRVDFK CYDEPHDFKS LPNLAIGSVK LLTVVDCVLD DDRPILESF KFLEVADVRS FVYNNHENGI RYNAKYFEGM EQLENLTLAR GVVSIDRDTF SGFLNLKRLT IEHNKLNLQP G TFEALSNL ...String:
TSTKRFTCPE ESEASNCSCE EFPSKTHFYC PDFNPTLYVD VEDRMRVDFK CYDEPHDFKS LPNLAIGSVK LLTVVDCVLD DDRPILESF KFLEVADVRS FVYNNHENGI RYNAKYFEGM EQLENLTLAR GVVSIDRDTF SGFLNLKRLT IEHNKLNLQP G TFEALSNL TYLGLVYNGL NEIQPGLFDG LESLEALSLS YNDIKSLSAG SFNGLSSLRM LNLRVNKIES FDANTFASLK EL SRLEITL NPFVSLPRGL FSENKKLKTL ILTNNRKLVT LPEELLANLK ELTVVNLSHN GVGNLPESLL SGSSGIIELN LGY NRLNSL PEELLSDQPQ LQVLNLDHNQ LESIPDYFLE RNVELQTLYL SHNRLRSLSE KAFTKLKNLK ELHLENNQLQ TIPQ FLFSG TPKLEEIYMQ NNQLALHANS FINEELSIAD NDNTPFQVLQ KLRILHLRNN SISTIFQDWY INNLEMQSLD LSFNK LPGL SYTQLQFQSN ITLNLSNNEI SQVLLIDDLD LQPYQRINVD LNHNPLNCNC NALKFIQLIQ SKAEHGLQFN VDQLRC SEP PNLLDATMDQ LQTKDLLCDF ESADDCPKDC QCAMRLLDHT VIVNCSGRGL TEFPDLPIPS QLHEDFNALE VHVENNR LT KLPNLTKHNE ITQLYARNNS IQNLLPHNIP SKLRIIDLSQ NLLKMIDDST LAQINRSSHL ETIRLSQNQW LCDCPASS F LIFVQQNSRL ISDMSAIRCH PSGKSLDSIT VNELCFEDYT TENLYFQ

UniProtKB: TIR domain-containing protein

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Macromolecule #2: AAEL013433-PA

MacromoleculeName: AAEL013433-PA / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Aedes aegypti (yellow fever mosquito)
Molecular weightTheoretical: 12.998096 KDa
Recombinant expressionOrganism: Baculovirus expression vector pFastBac1-HM
SequenceString:
GSDTANAPFL CESEQLLIHP KEELSRNNSM VWIVNTKDYK QGVRIEKCLK RQLGKPCNFC DADTECKQLF HYRTLVAVDK VTKKPYKEQ VLLPSCCKCA KILSTGWSHP QFEK

UniProtKB: AAEL013433-PA

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Macromolecule #5: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 5 / Number of copies: 3 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration5 mg/mL
BufferpH: 7.5
Component:
ConcentrationFormulaName
50.0 mMNaClsodium chloride
50.0 mMC4H11NO3Tris-HCl
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 12 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: OTHER
VitrificationCryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK I / Details: Blotting force 0.

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average exposure time: 2.0 sec. / Average electron dose: 51.1 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 51.1 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.0 mm / Nominal defocus max: 2.7 µm / Nominal defocus min: 1.0 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 507268
Startup modelType of model: INSILICO MODEL
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.74 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC / Number images used: 42866
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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