Key resources table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-FLAG M2 antibody | Sigma-Aldrich | Cat# F1804, RRID: AB_262044 |
| Goat polyclonal anti-Mouse IgG (H+L), HRP conjugated | Invitrogen | Cat# 62-6520, RRID: AB_2533947 |
| Bacterial and Virus Strains | ||
| Rosetta 2 (DE3) | Sigma-Aldrich | Cat# 71400-M |
| Chemicals, Peptides, and Recombinant Proteins | ||
| C-terminal protein C-6xHis tagged full-length EBV BILF1 | This paper | N/A |
| Gαi1-Gβ1-Gγ2 (GDP-bound) | Produced in house | N/A |
| scFv16 | Produced in house | N/A |
| 3C protease | Produced in house | N/A |
| Uranyl formate | VWR | Cat# 101410-83 |
| Lauryl Maltose Neopentyl Glycol (LMNG) | Anatrace | Cat# NG310 |
| n-Dodecyl-β-D-Maltoside (DDM) | Anatrace | Cat# D310 |
| glyco-diosgenin (GDN) | Anatrace | Cat# GDN101 |
| n-Octyl-β-D-Glucopyranoside (βOG) | Anatrace | Cat# O311 |
| Cholesteryl Hemisuccinate (CHS) | Anatrace | Cat# CH210 |
| Sodium cholate | Sigma-Aldrich | Cat# C6445 |
| Protease Inhibitor Cocktail | Sigma-Aldrich | Cat# P8849 |
| cOmplete Protease Inhibitor Cocktail | Roche | Cat# 05056489001 |
| HEPES | Fisher Scientific | Cat# BP310 |
| Tris Base | Fisher Scientific | Cat# BP152 |
| Tris Hydrochloride | Fisher Scientific | Cat# BP153 |
| Glycerol | Fisher Scientific | Cat# BP229 |
| Iodoacetamide | Sigma-Aldrich | Cat# I1149 |
| Sodium Chloride | Fisher Scientific | Cat# S271 |
| Calcium chloride dihydrate | Sigma-Aldrich | Cat# C7902 |
| Magnesium chloride hexahydrate | Sigma-Aldrich | Cat# M2393 |
| Manganese(II) chloride tetrahydrate | Sigma-Aldrich | Cat# M3634 |
| TCEP solution | Thermo Scientific | Cat# 77720 |
| EDTA | Fisher Scientific | Cat# BP120 |
| Protein C peptide | GenScript | N/A |
| Methyl α-D-mannopyranoside | Sigma-Aldrich | Cat# M6882 |
| Methyl α-D-glucopyranoside | Sigma-Aldrich | Cat# M9376 |
| 2-Mercaptoethanol | Sigma-Aldrich | Cat# M3148 |
| Imidazole | ACROS Organics | Cat# 122025000 |
| GDP | Sigma Aldrich | Cat# G7127 |
| Apyrase | New England Biolabs | Cat# M0398S |
| Antarctic Phosphatase | New England Biolabs | Cat# M0289S |
| Alkaline Phosphatase, Calf Intestinal | New England Biolabs | Cat# M0290S |
| Lambda Protein Phosphatase | New England Biolabs | Cat# P0753S |
| FreeStyle 293 Expression Medium | Thermo Fisher | Cat# 12338026 |
| ESF 921 Insect Cell Culture Medium, Protein Free | Expression Systems | Cat# 96-001-01 |
| Sf-900 III SFM (Serum-Free Media) | Thermo Fisher | Cat# 12658019 |
| GlutaMAX | Thermo Fisher | Cat# 35050079 |
| Fetal Bovine Serum (FBS) | Sigma-Aldrich | Cat# F7524 |
| Sodium butyrate | Sigma-Aldrich | Cat# 303410 |
| FuGENE HD Transfection Reagent | Promega | Cat# E2312 |
| Lipofectamine 2000 transfection reagent | Invitrogen | Cat# 11668019 |
| Dulbecco’s Modified Eagle Medium (DMEM) | Thermo Fisher | Cat# 10569010 |
| Penicillin solution | Immunology and Microbiology, Substrate Department, Faculty of Health and Medical Sciences, University of Copenhagen | N/A |
| Streptomycin solution | Immunology and Microbiology, Substrate Department, Faculty of Health and Medical Sciences, University of Copenhagen | N/A |
| Inositol, Myo, -[2-3H(N)]-, Specific Activity: 10-25Ci (370-925GBq)/mmol | Perkin Elmer | Cat# NET114A005MC |
| Hanks’ balanced salt solution (HBSS) | Thermo Fisher | Cat# 14025100 |
| YSi-poly-d-Lys-coated SPA beads | Perkin Elmer | Cat# RPNQ0010 |
| Poly-D-lysine | Thermo Fisher | Cat# A3890401 |
| Forskolin | Sigma-Aldrich | Cat# F6886 |
| Formaldehyde solution | Sigma-Aldrich | Cat# 252549 |
| Saponin from quillaja bark | Sigma-Aldrich | Cat# S7900 |
| 3, 3', 5, 5'-tetramethylbenzidine (TMB) substrate | Eco-tek | Cat# 4395A |
| Critical Commercial Assays | ||
| Steadylite plus reporter gene assay system | Perkin Elmer | Cat# 6066759 |
| Deposited Data | ||
| Cryo-EM structure of BILF1-Gi-scFv16 | RCSB PDB | PDB ID: 7JHJ |
| Cryo-EM map of BILF1-Gi-scFv16 | EMDB | EMD-22338 |
| Experimental Models: Cell Lines | ||
| Homo Sapiens: HEK293 cells | ATCC | CRL-1573 |
| Homo Sapiens: HEK293S GnTI− cells | Reeves, P.J. et al., Proc Natl Acad Sci USA. (2002), 99(21), 13419-13424. | CRL-3022 |
| Mus musculus: HPC-4 MOUSE HYBRIDOMA | ATCC | HB-9892 |
| Spodoptera frugiperda: Sf9 cells | ATCC | CRL-1711 |
| Trichuplusia ni: Tni or Hi5 cells | Expression Systems | Cat# 94-002 |
| Recombinant DNA | ||
| pVLAD6 full-length EBV BILF1 | This paper | N/A |
| pVL1392 human Gαi1 | Maeda, S. et al. Nat Commun. (2018), 9, 3712. | N/A |
| pVL1392duet human Gβ1-3C protease site-6xHis Gγ2 | Maeda, S. et al. Nat Commun. (2018), 9, 3712. | N/A |
| pVL1392 gp67 signal peptide-scFv16-3C protease site-6xHis | Maeda, S. et al. Nat Commun. (2018), 9, 3712. | N/A |
| BestBac 1.0 Linearized Baculovirus DNA | Expression Systems | Cat# 91-001 |
| pcDNA3.1+ | Genscript | Cat# SC1317 |
| pcDNA3.1+ GαΔ6qi4myr | Kostenis, E. et al. J Pharmacol Exp Ther. (2005). 313, 78-87. | N/A |
| pLuc-MCS CRE-cis reporter plasmid | Agilent-Stratagene | Cat# 219075 |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 | Paulsen, S. et al. J Virol (2005). 79, 536–46. | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring K122A mutation (K122A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring L167G N168G R169G mutation and G170 P171 N172 deletion (ECLGGG) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring L167E N168A R169D G170D P171R N172Y M173I mutation (ECLCXCR4a) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring R175D E176R G177F P178Y T179P K180N mutation (ECLCXCR4b) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring L167E N168A R169D G170D P171R N172Y M173I R175D E176R G177F P178Y T179P K180N mutation (ECLCXCR4ab) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring M240A L241A L243A Y267A mutation (TM6x7-4A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring M240A L241A L243A Y267A A271F mutation (TM6x7-4A1F) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring A271F mutation (A271F) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring M240F mutation (M240F) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring L241F mutation (L241F) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring Y282A mutation (Y282A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring E121A mutation (E121A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring H115A mutation (H115A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring L75A mutation (L75A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring L278N I279P mutation (NPxxY) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring S237C mutation (CWxM) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring S237C M240P mutation (CWxP) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring M240P mutation (SWxP) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring A125F mutation (A125F) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring H288A mutation (H288A) | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring F286A H288A mutation | This paper | N/A |
| pcDNA3.1+ N-terminal FLAG tagged full-length EBV BILF1 harboring H66A F286A H288A mutation | This paper | N/A |
| Software and Algorithms | ||
| SWISS-MODEL | Waterhouse, A. et al., Nucleic Acids Res. (2018). 46(W1), W296-W303 | https://swissmodel.expasy.org/ |
| SerialEM | Mastronarde, D.N. J. Struct. Biol. (2005). 152, 36-51. | http://bio3d.colorado.edu/SerialEM/ |
| MotionCor2 | Zheng S.Q. et al., Nat. Methods (2017). 14, 331-332. | http://msg.ucsf.edu/em/software/motioncor2.html |
| Relion 3.1 | Zivanov, J. et al. eLife (2018). e42166. | https://github.com/3dem/relion |
| Bsoft | Heymann, J.B. et al. Protein Sci. (2018). 27, 159-171. | https://lsbr.niams.nih.gov/bsoft/ |
| Coot 0.9 | Emsley P., Cowtan K. Acta Cryst. (2004). D60, 2126-2132. | https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ |
| Phenix suite | Adams, P.V. et al., Acta Cryst. (2010). D66, 213-221. | https://www.phenix-online.org/ |
| Isolde | Croll, T.I. Acta Cryst. (2018). D74, 519-530. | http://isoldehttps//isolde.cimr.cam.ac.uk/ |
| UCSF Chimera 1.14 | Pettersen, E.F. et al., J Comput Chem. (2004) 1605-1612. | https://www.cgl.ucsf.edu/chimera/ |
| UCSF ChimeraX 1.0 | Pettersen, E.F. et al., Protein Sci. (2020). | https://www.rbvi.ucsf.edu/chimerax/ |
| PyMol 2.3.0 | Schrödinger | http://www.pymol.org/;RRID: SCR_000305 |
| Prism 8 | GraphPad | https://www.graphpad.com/ |
| Other | ||
| Anti-Protein C affinity agarose | in-house | N/A |
| Agarose bound Concanavalin A | Vector Laboratories | Cat# AL-1003 |
| Ni-NTA Agarose | QIAGEN | Cat# 30250 |