Supramolecule #1: alpha4-beta3-delta GABA-A receptor in complex with DS2-Me, GABA, ...
Supramolecule
Name: alpha4-beta3-delta GABA-A receptor in complex with DS2-Me, GABA, nanobody Nb24 and megabody Mb30 type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#5 Details: The heteropentameric alpha4beta3delta GABA(A) receptor was recombinantly expressed in Expi293 cells. Constructs were engineered with the M3- M4 intracellular domain replaced either by a GLVI ...Details: The heteropentameric alpha4beta3delta GABA(A) receptor was recombinantly expressed in Expi293 cells. Constructs were engineered with the M3- M4 intracellular domain replaced either by a GLVI linker sequence (SQPARAA) or by a modified GLVI sequence containing Escherichia coli cytochrome B562RIL41. The sample was prepared in the presence of DS2-Me, GABA, nanobody Nb24, and megabody Mb30.
Name: Gamma-aminobutyric acid receptor subunit beta-3,Soluble cytochrome b562 type: protein_or_peptide / ID: 2 Details: Modified glvi sequence containing the Escherichia coli soluble cytochrome B562RIL41 (BRIL, amino acids 23-130, ADLE...QKYL, Uniprot P0ABE7) to give the sequence SQPAGT-BRIL-TGRAA. Linker ...Details: Modified glvi sequence containing the Escherichia coli soluble cytochrome B562RIL41 (BRIL, amino acids 23-130, ADLE...QKYL, Uniprot P0ABE7) to give the sequence SQPAGT-BRIL-TGRAA. Linker regions substituted Gly308-Asn421. This BRIL domain was present in the protein but not resolved in the cryo-EM density and is therefore not included in the atomic model,Modified glvi sequence containing the Escherichia coli soluble cytochrome B562RIL41 (BRIL, amino acids 23-130, ADLE...QKYL, Uniprot P0ABE7) to give the sequence SQPAGT-BRIL-TGRAA. Linker regions substituted Gly308-Asn421. This BRIL domain was present in the protein but not resolved in the cryo-EM density and is therefore not included in the atomic model,Modified glvi sequence containing the Escherichia coli soluble cytochrome B562RIL41 (BRIL, amino acids 23-130, ADLE...QKYL, Uniprot P0ABE7) to give the sequence SQPAGT-BRIL-TGRAA. Linker regions substituted Gly308-Asn421. This BRIL domain was present in the protein but not resolved in the cryo-EM density and is therefore not included in the atomic model,Modified glvi sequence containing the Escherichia coli soluble cytochrome B562RIL41 (BRIL, amino acids 23-130, ADLE...QKYL, Uniprot P0ABE7) to give the sequence SQPAGT-BRIL-TGRAA. Linker regions substituted Gly308-Asn421. This BRIL domain was present in the protein but not resolved in the cryo-EM density and is therefore not included in the atomic model Number of copies: 2 / Enantiomer: LEVO
Name: Gamma-aminobutyric acid receptor subunit delta / type: protein_or_peptide / ID: 3 Details: Includes an SBP tag at start of the sequence, this is not resolved in density and not modelled. Glvi linker sequence (SQPARAA) substituted between Asp312-Asp401.,Includes an SBP tag at start ...Details: Includes an SBP tag at start of the sequence, this is not resolved in density and not modelled. Glvi linker sequence (SQPARAA) substituted between Asp312-Asp401.,Includes an SBP tag at start of the sequence, this is not resolved in density and not modelled. Glvi linker sequence (SQPARAA) substituted between Asp312-Asp401.,Includes an SBP tag at start of the sequence, this is not resolved in density and not modelled. Glvi linker sequence (SQPARAA) substituted between Asp312-Asp401.,Includes an SBP tag at start of the sequence, this is not resolved in density and not modelled. Glvi linker sequence (SQPARAA) substituted between Asp312-Asp401. Number of copies: 1 / Enantiomer: LEVO
Name: Megabody Mb30 / type: protein_or_peptide / ID: 4 Details: This nanobody was expressed as part of a megabody fusion construct. The scaffold domain of the megabody was present in the construct but was disordered in the cryo-EM density and therefore ...Details: This nanobody was expressed as part of a megabody fusion construct. The scaffold domain of the megabody was present in the construct but was disordered in the cryo-EM density and therefore not included in the atomic model. Only the nanobody sequence is provided in the deposited coordinates. Number of copies: 2 / Enantiomer: LEVO
Type of model: NONE Details: An ab initio 3D reconstruction was generated from experimental 2D class averages in cryoSPARC without the use of any external reference, and was used as the startup model for future refinements.
Final reconstruction
Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.04 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.4.1 to 4.7.1) / Number images used: 110542
Initial angle assignment
Type: PROJECTION MATCHING / Software - Name: cryoSPARC (ver. 4.4.1 to 4.7.1) Details: Particle orientations were assigned by projection matching during iterative refinement in cryoSPARC.
Final angle assignment
Type: PROJECTION MATCHING / Software - Name: cryoSPARC (ver. 4.4.1 to 4.7.1) Details: Final orientations determined by projection matching during non-uniform refinement in cryoSPARC.
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