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- PDB-9pxb: Hamster Scap/Insig-2 complex with cholesterol and bound Fab4G10 -

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Basic information

Entry
Database: PDB / ID: 9pxb
TitleHamster Scap/Insig-2 complex with cholesterol and bound Fab4G10
Components
  • Fab4G10 heavy chain
  • Fab4G10 light chain
  • Insulin-induced gene protein
  • Sterol regulatory element-binding protein cleavage-activating protein
KeywordsMEMBRANE PROTEIN / Complex / sterol-binding / COPII-binding
Function / homology
Function and homology information


SREBP-SCAP complex retention in endoplasmic reticulum / vesicle coat / SREBP-SCAP-Insig complex / SREBP-SCAP complex / regulation of cholesterol biosynthetic process / sterol binding / SREBP signaling pathway / COPII-coated vesicle cargo loading / cellular response to sterol / cholesterol biosynthetic process ...SREBP-SCAP complex retention in endoplasmic reticulum / vesicle coat / SREBP-SCAP-Insig complex / SREBP-SCAP complex / regulation of cholesterol biosynthetic process / sterol binding / SREBP signaling pathway / COPII-coated vesicle cargo loading / cellular response to sterol / cholesterol biosynthetic process / cholesterol metabolic process / ER to Golgi transport vesicle membrane / cellular response to insulin stimulus / protein import into nucleus / Golgi membrane / lipid binding / endoplasmic reticulum membrane / endoplasmic reticulum / membrane
Similarity search - Function
Insulin-induced protein family / Insulin-induced protein (INSIG) / : / : / SCAP N-terminal / SCAP Beta-propeller / Sterol regulatory element-binding protein cleavage-activating protein / : / Sterol-sensing domain of SREBP cleavage-activation / Sterol-sensing domain (SSD) profile. ...Insulin-induced protein family / Insulin-induced protein (INSIG) / : / : / SCAP N-terminal / SCAP Beta-propeller / Sterol regulatory element-binding protein cleavage-activating protein / : / Sterol-sensing domain of SREBP cleavage-activation / Sterol-sensing domain (SSD) profile. / Sterol-sensing domain / WD40 repeat, conserved site / Trp-Asp (WD) repeats signature. / Trp-Asp (WD) repeats profile. / Trp-Asp (WD) repeats circular profile. / WD40 repeats / WD40 repeat / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
Similarity search - Domain/homology
CHOLESTEROL / Sterol regulatory element-binding protein cleavage-activating protein / Insulin-induced gene protein
Similarity search - Component
Biological speciesCricetulus griseus (Chinese hamster)
Mus musculus (house mouse)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsWilliams, B.C. / Kober, D.L. / Bai, X. / Radhakrishnan, A. / Rosenbaum, D.M.
Funding support United States, 3items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35GM116387 United States
National Institutes of Health/National Heart, Lung, and Blood Institute (NIH/NHLBI)P01HL160487 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)R01AI158357 United States
CitationJournal: To Be Published
Title: Hamster Scap/Insig-2 bound to cholesterol
Authors: Williams, B.C. / Rosenbaum, D.M.
History
DepositionAug 5, 2025Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 24, 2025Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: EM metadata / Data content type: EM metadata / Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: Additional map / Part number: 1 / Data content type: Additional map / Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: FSC / Data content type: FSC / Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: Half map / Part number: 1 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: Half map / Part number: 2 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: Image / Data content type: Image / Provider: repository / Type: Initial release
Revision 1.0Dec 24, 2025Data content type: Primary map / Data content type: Primary map / Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Sterol regulatory element-binding protein cleavage-activating protein
C: Insulin-induced gene protein
H: Fab4G10 heavy chain
L: Fab4G10 light chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)168,0918
Polymers166,5074
Non-polymers1,5844
Water00
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1

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Components

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Protein , 2 types, 2 molecules AC

#1: Protein Sterol regulatory element-binding protein cleavage-activating protein / SCAP / SREBP cleavage-activating protein


Mass: 89014.695 Da / Num. of mol.: 1 / Mutation: V133A, T140V, H244D, A245S, K246R, A253S, M256K
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Cricetulus griseus (Chinese hamster) / Gene: SCAP / Production host: Homo sapiens (human) / References: UniProt: P97260
#2: Protein Insulin-induced gene protein


Mass: 24929.936 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Cricetulus griseus (Chinese hamster) / Gene: Insig2 / Production host: Homo sapiens (human) / References: UniProt: Q8CFA5

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Antibody , 2 types, 2 molecules HL

#3: Antibody Fab4G10 heavy chain


Mass: 27016.852 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Production host: Homo sapiens (human)
#4: Antibody Fab4G10 light chain


Mass: 25545.189 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Production host: Homo sapiens (human)

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Sugars / Non-polymers , 2 types, 4 molecules

#5: Polysaccharide 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose


Type: oligosaccharide / Mass: 424.401 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
DescriptorTypeProgram
DGlcpNAcb1-4DGlcpNAcb1-Glycam Condensed SequenceGMML 1.0
WURCS=2.0/1,2,1/[a2122h-1b_1-5_2*NCC/3=O]/1-1/a4-b1WURCSPDB2Glycan 1.1.0
[]{[(4+1)][b-D-GlcpNAc]{[(4+1)][b-D-GlcpNAc]{}}}LINUCSPDB-CARE
#6: Chemical ChemComp-CLR / CHOLESTEROL


Mass: 386.654 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C27H46O / Feature type: SUBJECT OF INVESTIGATION

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Details

Has ligand of interestY
Has protein modificationY

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

Component
IDNameTypeEntity IDParent-IDSource
1Scap/Insig-2 complex with bound Fab4G10COMPLEX#1-#40RECOMBINANT
2Scap/Insig-2 complexCOMPLEX#1-#21RECOMBINANT
3Fab4G10COMPLEX#3-#41RECOMBINANT
Source (natural)
IDEntity assembly-IDOrganismNcbi tax-ID
21Cricetulus griseus (Chinese hamster)10029
32Cricetulus griseus (Chinese hamster)10029
43Mus musculus (house mouse)10090
Source (recombinant)
IDEntity assembly-IDOrganismNcbi tax-ID
21Homo sapiens (human)9606
32Homo sapiens (human)9606
43Homo sapiens (human)9606
Buffer solutionpH: 7.5
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: TFS KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal defocus max: 2400 nm / Nominal defocus min: 1400 nm
Image recordingElectron dose: 60 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k)

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Processing

EM software
IDNameVersionCategory
1RELIONparticle selection
2PHENIX1.20.1_4487:model refinement
13RELION3D reconstruction
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
3D reconstructionResolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 160849 / Symmetry type: POINT
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.0027842
ELECTRON MICROSCOPYf_angle_d0.51810706
ELECTRON MICROSCOPYf_dihedral_angle_d5.7481071
ELECTRON MICROSCOPYf_chiral_restr0.0391238
ELECTRON MICROSCOPYf_plane_restr0.0041325

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