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- EMDB-13596: Structure of thermostabilised human NTCP in complex with Megabody 91 -

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Basic information

Entry
Database: EMDB / ID: EMD-13596
TitleStructure of thermostabilised human NTCP in complex with Megabody 91
Map data
Sample
  • Complex: human NTCP complexed with Megabody 91
    • Protein or peptide: Sodium/bile acid cotransporter
    • Protein or peptide: Anti-RON nanobody,Megabody 91,Glucosidase YgjK
Function / homology
Function and homology information


glucosidase complex / alpha,alpha-trehalase activity / bile acid:sodium symporter activity / trehalose catabolic process / glucosidase activity / bile acid signaling pathway / regulation of bile acid secretion / oligosaccharide catabolic process / Hydrolases; Glycosylases; Glycosidases, i.e. enzymes that hydrolyse O- and S-glycosyl compounds / bile acid and bile salt transport ...glucosidase complex / alpha,alpha-trehalase activity / bile acid:sodium symporter activity / trehalose catabolic process / glucosidase activity / bile acid signaling pathway / regulation of bile acid secretion / oligosaccharide catabolic process / Hydrolases; Glycosylases; Glycosidases, i.e. enzymes that hydrolyse O- and S-glycosyl compounds / bile acid and bile salt transport / Recycling of bile acids and salts / response to nutrient levels / response to organic cyclic compound / response to estrogen / cellular response to xenobiotic stimulus / virus receptor activity / basolateral plasma membrane / response to ethanol / DNA damage response / metal ion binding / plasma membrane
Similarity search - Function
: / Glucosidase YgjK, N-terminal / Glycoside hydrolase, family 37 / Mannosylglycerate hydrolase MGH1-like glycoside hydrolase domain / Bile acid:sodium symporter/arsenical resistance protein Acr3 / Bile acid:sodium symporter / Sodium Bile acid symporter family / Sodium/solute symporter superfamily / Six-hairpin glycosidase-like superfamily / Six-hairpin glycosidase superfamily
Similarity search - Domain/homology
Glucosidase YgjK / Hepatic sodium/bile acid cotransporter
Similarity search - Component
Biological speciesHomo sapiens (human) / Lama glama (llama)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.3 Å
AuthorsGoutam K / Reyes N
Funding supportEuropean Union, 1 items
OrganizationGrant numberCountry
European Research Council (ERC)309657European Union
CitationJournal: Nature / Year: 2022
Title: Structural basis of sodium-dependent bile salt uptake into the liver.
Authors: Kapil Goutam / Francesco S Ielasi / Els Pardon / Jan Steyaert / Nicolas Reyes /
Abstract: The liver takes up bile salts from blood to generate bile, enabling absorption of lipophilic nutrients and excretion of metabolites and drugs. Human Na-taurocholate co-transporting polypeptide (NTCP) ...The liver takes up bile salts from blood to generate bile, enabling absorption of lipophilic nutrients and excretion of metabolites and drugs. Human Na-taurocholate co-transporting polypeptide (NTCP) is the main bile salt uptake system in liver. NTCP is also the cellular entry receptor of human hepatitis B and D viruses (HBV/HDV), and has emerged as an important target for antiviral drugs. However, the molecular mechanisms underlying NTCP transport and viral receptor functions remain incompletely understood. Here we present cryo-electron microscopy structures of human NTCP in complexes with nanobodies, revealing key conformations of its transport cycle. NTCP undergoes a conformational transition opening a wide transmembrane pore that serves as the transport pathway for bile salts, and exposes key determinant residues for HBV/HDV binding to the outside of the cell. A nanobody that stabilizes pore closure and inward-facing states impairs recognition of the HBV/HDV receptor-binding domain preS1, demonstrating binding selectivity of the viruses for open-to-outside over inward-facing conformations of the NTCP transport cycle. These results provide molecular insights into NTCP 'gated-pore' transport and HBV/HDV receptor recognition mechanisms, and are expected to help with development of liver disease therapies targeting NTCP.
History
DepositionSep 18, 2021-
Header (metadata) releaseMay 18, 2022-
Map releaseMay 18, 2022-
UpdateJul 6, 2022-
Current statusJul 6, 2022Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_13596.png

Downloads & links

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Map

FileDownload / File: emd_13596.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.01 Å/pix.
x 280 pix.
= 282.24 Å		1.01 Å/pix.
x 280 pix.
= 282.24 Å		1.01 Å/pix.
x 280 pix.
= 282.24 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.008 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.1
Minimum - Maximum-0.648644 - 2.3310084
Average (Standard dev.)0.000469606 (±0.015360326)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions280280280
Spacing280280280
CellA=B=C: 282.24 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : human NTCP complexed with Megabody 91

EntireName: human NTCP complexed with Megabody 91
Components
  • Complex: human NTCP complexed with Megabody 91
    • Protein or peptide: Sodium/bile acid cotransporter
    • Protein or peptide: Anti-RON nanobody,Megabody 91,Glucosidase YgjK

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Supramolecule #1: human NTCP complexed with Megabody 91

SupramoleculeName: human NTCP complexed with Megabody 91 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: Sodium/bile acid cotransporter

MacromoleculeName: Sodium/bile acid cotransporter / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 36.652391 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MAHTASAPFT FTLPPNFGKR PTDLALSVIL VVMLFIIMLS LGCTMEFSKI KAHLWKPKGL AIALVAQYGI MPLTAFVLGK VFRLNNIEA LAILICGCSP GGNLSNIFSL AMKGDMNLSI VMTTCSTFLA LGMMPLLLYI YSRGIYDGDL KDKVPYKGIV I SLVLVLIP ...String:
MAHTASAPFT FTLPPNFGKR PTDLALSVIL VVMLFIIMLS LGCTMEFSKI KAHLWKPKGL AIALVAQYGI MPLTAFVLGK VFRLNNIEA LAILICGCSP GGNLSNIFSL AMKGDMNLSI VMTTCSTFLA LGMMPLLLYI YSRGIYDGDL KDKVPYKGIV I SLVLVLIP CTIGIVLKSK RPQYMRYVIK GGMIIILLCS VAVTVLSAIN VGKSIMFAMT PHLIATSSLM PFIGFLLGYV LS ALFCLNG RCRRTVSMET GCQNVQLCST ILNVAFPPEV IGPLFFFPLL YMIFQLGEGL LLIAIFWCYE KFKTPKDKTK MIY TAATTE ELEVLFQ

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Macromolecule #2: Anti-RON nanobody,Megabody 91,Glucosidase YgjK

MacromoleculeName: Anti-RON nanobody,Megabody 91,Glucosidase YgjK / type: protein_or_peptide / ID: 2
Details: The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The ...Details: The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The alignment software here is misaligning N-terminal part so I have just put build sequence in the sample sequence box. Here is the full sequence: QVQLVESGGGLV KEETQSGLNNYARVVEKGQYDSLEIPAQVAASWESGRDDAAVFGFIDKEQLDKYVANGGKRSDWTVKFAENRSQDGTLLGYSLLQESVDQASYMYSDNHYLAEMATILGKPEEAKRYRQLAQQLADYINTCMFDPTTQFYYDVRIEDKPLANGCAGKPIVERGKGPEGWSPLFNGAATQANADAVVKVMLDPKEFNTFVPLGTAALTNPAFGADIYWRGRVWVDQFWFGLKGMERYGYRDDALKLADTFFRHAKGLTADGPIQENYNPLTGAQQGAPNFSWSAAHLYMLYNDFFRKQASGGGSGGGGSGGGGSGNADNYKNVINRTGAPQYMKDYDYDDHQRFNPFFDLGAWHGHLLPDGPNTMGGFPGVALLTEEYINFMASNFDRLTVWQDGKKVDFTLEAYSIPGALVQKLTAKDVQVEMTLRFATPRTSLLETKITSNKPLDLVWDGELLEKLEAKEGKPLSDKTIAGEYPDYQRKISATRDGLKVTFGKVRATWDLLTSGESEYQVHKSLPVQTEINGNRFTSKAHINGSTTLYTTYSHLLTAQEVSKEQMQIRDILARPAFYLTASQQRWEEYLKKGLTNPDATPEQTRVAVKAIETLNGNWRSPGGAVKFNTVTPSVTGRWFSGNQTWPWDTWKQAFAMAHFNPDIAKENIRAVFSWQIQPGDSVRPQDVGFVPDLIAWNLSPERGGDGGNWNERNTKPSLAAWSVMEVYNVTQDKTWVAEMYPKLVAYHDWWLRNRDHNGNGVPEYGATRDKAHNTESGEMLFTVKK SLRLSCAASTNLRSYAMAWFRQAPGKEREFVSFINWNYGNTRYA DSVKGRFTISRDNAKITVYLQMNSLKPEDTAVYYCAAATIGRLAGIDSTTLYDYWGQGTQ VTVSSHHHHHHEPEA,The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The alignment software here is misaligning N-terminal part so I have just put build sequence in the sample sequence box. Here is the full sequence: QVQLVESGGGLV KEETQSGLNNYARVVEKGQYDSLEIPAQVAASWESGRDDAAVFGFIDKEQLDKYVANGGKRSDWTVKFAENRSQDGTLLGYSLLQESVDQASYMYSDNHYLAEMATILGKPEEAKRYRQLAQQLADYINTCMFDPTTQFYYDVRIEDKPLANGCAGKPIVERGKGPEGWSPLFNGAATQANADAVVKVMLDPKEFNTFVPLGTAALTNPAFGADIYWRGRVWVDQFWFGLKGMERYGYRDDALKLADTFFRHAKGLTADGPIQENYNPLTGAQQGAPNFSWSAAHLYMLYNDFFRKQASGGGSGGGGSGGGGSGNADNYKNVINRTGAPQYMKDYDYDDHQRFNPFFDLGAWHGHLLPDGPNTMGGFPGVALLTEEYINFMASNFDRLTVWQDGKKVDFTLEAYSIPGALVQKLTAKDVQVEMTLRFATPRTSLLETKITSNKPLDLVWDGELLEKLEAKEGKPLSDKTIAGEYPDYQRKISATRDGLKVTFGKVRATWDLLTSGESEYQVHKSLPVQTEINGNRFTSKAHINGSTTLYTTYSHLLTAQEVSKEQMQIRDILARPAFYLTASQQRWEEYLKKGLTNPDATPEQTRVAVKAIETLNGNWRSPGGAVKFNTVTPSVTGRWFSGNQTWPWDTWKQAFAMAHFNPDIAKENIRAVFSWQIQPGDSVRPQDVGFVPDLIAWNLSPERGGDGGNWNERNTKPSLAAWSVMEVYNVTQDKTWVAEMYPKLVAYHDWWLRNRDHNGNGVPEYGATRDKAHNTESGEMLFTVKK SLRLSCAASTNLRSYAMAWFRQAPGKEREFVSFINWNYGNTRYA DSVKGRFTISRDNAKITVYLQMNSLKPEDTAVYYCAAATIGRLAGIDSTTLYDYWGQGTQ VTVSSHHHHHHEPEA,The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The alignment software here is misaligning N-terminal part so I have just put build sequence in the sample sequence box. Here is the full sequence: QVQLVESGGGLV KEETQSGLNNYARVVEKGQYDSLEIPAQVAASWESGRDDAAVFGFIDKEQLDKYVANGGKRSDWTVKFAENRSQDGTLLGYSLLQESVDQASYMYSDNHYLAEMATILGKPEEAKRYRQLAQQLADYINTCMFDPTTQFYYDVRIEDKPLANGCAGKPIVERGKGPEGWSPLFNGAATQANADAVVKVMLDPKEFNTFVPLGTAALTNPAFGADIYWRGRVWVDQFWFGLKGMERYGYRDDALKLADTFFRHAKGLTADGPIQENYNPLTGAQQGAPNFSWSAAHLYMLYNDFFRKQASGGGSGGGGSGGGGSGNADNYKNVINRTGAPQYMKDYDYDDHQRFNPFFDLGAWHGHLLPDGPNTMGGFPGVALLTEEYINFMASNFDRLTVWQDGKKVDFTLEAYSIPGALVQKLTAKDVQVEMTLRFATPRTSLLETKITSNKPLDLVWDGELLEKLEAKEGKPLSDKTIAGEYPDYQRKISATRDGLKVTFGKVRATWDLLTSGESEYQVHKSLPVQTEINGNRFTSKAHINGSTTLYTTYSHLLTAQEVSKEQMQIRDILARPAFYLTASQQRWEEYLKKGLTNPDATPEQTRVAVKAIETLNGNWRSPGGAVKFNTVTPSVTGRWFSGNQTWPWDTWKQAFAMAHFNPDIAKENIRAVFSWQIQPGDSVRPQDVGFVPDLIAWNLSPERGGDGGNWNERNTKPSLAAWSVMEVYNVTQDKTWVAEMYPKLVAYHDWWLRNRDHNGNGVPEYGATRDKAHNTESGEMLFTVKK SLRLSCAASTNLRSYAMAWFRQAPGKEREFVSFINWNYGNTRYA DSVKGRFTISRDNAKITVYLQMNSLKPEDTAVYYCAAATIGRLAGIDSTTLYDYWGQGTQ VTVSSHHHHHHEPEA,The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The alignment software here is misaligning N-terminal part so I have just put build sequence in the sample sequence box. Here is the full sequence: QVQLVESGGGLV KEETQSGLNNYARVVEKGQYDSLEIPAQVAASWESGRDDAAVFGFIDKEQLDKYVANGGKRSDWTVKFAENRSQDGTLLGYSLLQESVDQASYMYSDNHYLAEMATILGKPEEAKRYRQLAQQLADYINTCMFDPTTQFYYDVRIEDKPLANGCAGKPIVERGKGPEGWSPLFNGAATQANADAVVKVMLDPKEFNTFVPLGTAALTNPAFGADIYWRGRVWVDQFWFGLKGMERYGYRDDALKLADTFFRHAKGLTADGPIQENYNPLTGAQQGAPNFSWSAAHLYMLYNDFFRKQASGGGSGGGGSGGGGSGNADNYKNVINRTGAPQYMKDYDYDDHQRFNPFFDLGAWHGHLLPDGPNTMGGFPGVALLTEEYINFMASNFDRLTVWQDGKKVDFTLEAYSIPGALVQKLTAKDVQVEMTLRFATPRTSLLETKITSNKPLDLVWDGELLEKLEAKEGKPLSDKTIAGEYPDYQRKISATRDGLKVTFGKVRATWDLLTSGESEYQVHKSLPVQTEINGNRFTSKAHINGSTTLYTTYSHLLTAQEVSKEQMQIRDILARPAFYLTASQQRWEEYLKKGLTNPDATPEQTRVAVKAIETLNGNWRSPGGAVKFNTVTPSVTGRWFSGNQTWPWDTWKQAFAMAHFNPDIAKENIRAVFSWQIQPGDSVRPQDVGFVPDLIAWNLSPERGGDGGNWNERNTKPSLAAWSVMEVYNVTQDKTWVAEMYPKLVAYHDWWLRNRDHNGNGVPEYGATRDKAHNTESGEMLFTVKK SLRLSCAASTNLRSYAMAWFRQAPGKEREFVSFINWNYGNTRYA DSVKGRFTISRDNAKITVYLQMNSLKPEDTAVYYCAAATIGRLAGIDSTTLYDYWGQGTQ VTVSSHHHHHHEPEA,The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The alignment software here is misaligning N-terminal part so I have just put build sequence in the sample sequence box. Here is the full sequence: QVQLVESGGGLV KEETQSGLNNYARVVEKGQYDSLEIPAQVAASWESGRDDAAVFGFIDKEQLDKYVANGGKRSDWTVKFAENRSQDGTLLGYSLLQESVDQASYMYSDNHYLAEMATILGKPEEAKRYRQLAQQLADYINTCMFDPTTQFYYDVRIEDKPLANGCAGKPIVERGKGPEGWSPLFNGAATQANADAVVKVMLDPKEFNTFVPLGTAALTNPAFGADIYWRGRVWVDQFWFGLKGMERYGYRDDALKLADTFFRHAKGLTADGPIQENYNPLTGAQQGAPNFSWSAAHLYMLYNDFFRKQASGGGSGGGGSGGGGSGNADNYKNVINRTGAPQYMKDYDYDDHQRFNPFFDLGAWHGHLLPDGPNTMGGFPGVALLTEEYINFMASNFDRLTVWQDGKKVDFTLEAYSIPGALVQKLTAKDVQVEMTLRFATPRTSLLETKITSNKPLDLVWDGELLEKLEAKEGKPLSDKTIAGEYPDYQRKISATRDGLKVTFGKVRATWDLLTSGESEYQVHKSLPVQTEINGNRFTSKAHINGSTTLYTTYSHLLTAQEVSKEQMQIRDILARPAFYLTASQQRWEEYLKKGLTNPDATPEQTRVAVKAIETLNGNWRSPGGAVKFNTVTPSVTGRWFSGNQTWPWDTWKQAFAMAHFNPDIAKENIRAVFSWQIQPGDSVRPQDVGFVPDLIAWNLSPERGGDGGNWNERNTKPSLAAWSVMEVYNVTQDKTWVAEMYPKLVAYHDWWLRNRDHNGNGVPEYGATRDKAHNTESGEMLFTVKK SLRLSCAASTNLRSYAMAWFRQAPGKEREFVSFINWNYGNTRYA DSVKGRFTISRDNAKITVYLQMNSLKPEDTAVYYCAAATIGRLAGIDSTTLYDYWGQGTQ VTVSSHHHHHHEPEA,The megabody scaffold is excluded from the structure during refinement and model building. The build model corresponds to residues 2-12 and 788-895 belonging to nanobody part only. The alignment software here is misaligning N-terminal part so I have just put build sequence in the sample sequence box. Here is the full sequence: QVQLVESGGGLV KEETQSGLNNYARVVEKGQYDSLEIPAQVAASWESGRDDAAVFGFIDKEQLDKYVANGGKRSDWTVKFAENRSQDGTLLGYSLLQESVDQASYMYSDNHYLAEMATILGKPEEAKRYRQLAQQLADYINTCMFDPTTQFYYDVRIEDKPLANGCAGKPIVERGKGPEGWSPLFNGAATQANADAVVKVMLDPKEFNTFVPLGTAALTNPAFGADIYWRGRVWVDQFWFGLKGMERYGYRDDALKLADTFFRHAKGLTADGPIQENYNPLTGAQQGAPNFSWSAAHLYMLYNDFFRKQASGGGSGGGGSGGGGSGNADNYKNVINRTGAPQYMKDYDYDDHQRFNPFFDLGAWHGHLLPDGPNTMGGFPGVALLTEEYINFMASNFDRLTVWQDGKKVDFTLEAYSIPGALVQKLTAKDVQVEMTLRFATPRTSLLETKITSNKPLDLVWDGELLEKLEAKEGKPLSDKTIAGEYPDYQRKISATRDGLKVTFGKVRATWDLLTSGESEYQVHKSLPVQTEINGNRFTSKAHINGSTTLYTTYSHLLTAQEVSKEQMQIRDILARPAFYLTASQQRWEEYLKKGLTNPDATPEQTRVAVKAIETLNGNWRSPGGAVKFNTVTPSVTGRWFSGNQTWPWDTWKQAFAMAHFNPDIAKENIRAVFSWQIQPGDSVRPQDVGFVPDLIAWNLSPERGGDGGNWNERNTKPSLAAWSVMEVYNVTQDKTWVAEMYPKLVAYHDWWLRNRDHNGNGVPEYGATRDKAHNTESGEMLFTVKK SLRLSCAASTNLRSYAMAWFRQAPGKEREFVSFINWNYGNTRYA DSVKGRFTISRDNAKITVYLQMNSLKPEDTAVYYCAAATIGRLAGIDSTTLYDYWGQGTQ VTVSSHHHHHHEPEA
Number of copies: 1 / Enantiomer: LEVO
EC number: Hydrolases; Glycosylases; Glycosidases, i.e. enzymes that hydrolyse O- and S-glycosyl compounds
Source (natural)Organism: Lama glama (llama)
Molecular weightTheoretical: 101.583031 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: QVQLVESGGG LVKEETQSGL NNYARVVEKG QYDSLEIPAQ VAASWESGRD DAAVFGFIDK EQLDKYVANG GKRSDWTVKF AENRSQDGT LLGYSLLQES VDQASYMYSD NHYLAEMATI LGKPEEAKRY RQLAQQLADY INTCMFDPTT QFYYDVRIED K PLANGCAG ...String:
QVQLVESGGG LVKEETQSGL NNYARVVEKG QYDSLEIPAQ VAASWESGRD DAAVFGFIDK EQLDKYVANG GKRSDWTVKF AENRSQDGT LLGYSLLQES VDQASYMYSD NHYLAEMATI LGKPEEAKRY RQLAQQLADY INTCMFDPTT QFYYDVRIED K PLANGCAG KPIVERGKGP EGWSPLFNGA ATQANADAVV KVMLDPKEFN TFVPLGTAAL TNPAFGADIY WRGRVWVDQF WF GLKGMER YGYRDDALKL ADTFFRHAKG LTADGPIQEN YNPLTGAQQG APNFSWSAAH LYMLYNDFFR KQASGGGSGG GGS GGGGSG NADNYKNVIN RTGAPQYMKD YDYDDHQRFN PFFDLGAWHG HLLPDGPNTM GGFPGVALLT EEYINFMASN FDRL TVWQD GKKVDFTLEA YSIPGALVQK LTAKDVQVEM TLRFATPRTS LLETKITSNK PLDLVWDGEL LEKLEAKEGK PLSDK TIAG EYPDYQRKIS ATRDGLKVTF GKVRATWDLL TSGESEYQVH KSLPVQTEIN GNRFTSKAHI NGSTTLYTTY SHLLTA QEV SKEQMQIRDI LARPAFYLTA SQQRWEEYLK KGLTNPDATP EQTRVAVKAI ETLNGNWRSP GGAVKFNTVT PSVTGRW FS GNQTWPWDTW KQAFAMAHFN PDIAKENIRA VFSWQIQPGD SVRPQDVGFV PDLIAWNLSP ERGGDGGNWN ERNTKPSL A AWSVMEVYNV TQDKTWVAEM YPKLVAYHDW WLRNRDHNGN GVPEYGATRD KAHNTESGEM LFTVKKSLRL SCAASTNLR SYAMAWFRQA PGKEREFVSF INWNYGNTRY ADSVKGRFTI SRDNAKITVY LQMNSLKPED TAVYYCAAAT IGRLAGIDST TLYDYWGQG TQVTVSSHHH HHHEPEA

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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

BufferpH: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 56.5 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: INSILICO MODEL
In silico model: The initial model for Nanobody was created using I-TASSER server.
Details: Transporter was modelled de novo.
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 184768
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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