REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Monoclonal ANTI-FLAG M2-HRP antibody	Sigma-Aldrich	Cat# A8592; RRID: AB_439702
Chemicals, peptides, and recombinant proteins
TRIS	SRL	Cat# 71033
HEPES	SRL	Cat# 63732
NaCl	SRL	Cat# 41721
EDTA	SRL	Cat# 12070
Phenylmethanesulfonyl Fluoride (PMSF)	SRL	Cat# 84375 (84375)
Benzamidine Hydrochloride	SRL	Cat# 93014 (0248255)
Lysozyme	SRL	Cat# 45822
Glycerol	SRL	Cat# 77453
Dithiothreitol	HiMedia	Cat# MB070
Lauryl Maltose Neopentyl Glycol (MNG)	Anatrace	Cat# NG310, CAS no.1257852-96-2
Paraformaldehyde (PFA)	Sigma Aldrich	Cat# P6148, CAS no. 30525-89-4
Poly-D-lysine	Sigma Aldrich	Cat# P0899
TMB (Tetramethylbenzidine)	Thermo Fisher Scientific	Cat# 34028
Janus Green B	Sigma Aldrich	Cat# 201677
PEI (Polyethylenimine)	Polysciences	Cat# 23966
Bovine Serum Albumin, BSA	SRL	Cat# 83803 (0140105)
HBSS - Hank’s Balanced Salt Solution	Thermo Fisher Scientific	Cat# 14065
GIBCO Fetal Bovine Serum	Thermo Fisher Scientific	Cat# 10270-106
DMEM	Cellclone	Cat# CC3004
Phosphate-buffered saline (PBS)	Sigma Aldrich	Cat# D1283
GIBCO Penicillin-Streptomycin	Thermo Fisher Scientific	Cat# 15140122
Coelenterazine	Goldbio	Cat# CZ05
Glyco-diosgenin (GDN)	Anatrace	GDN101
Cholesteryl Hemisuccinate	Sigma	C6512
Coomassie briliiant Blue	SRL	Cat# 64222
Uranyl formate	Polysciences	Cat# 24762-1
Recombinant rat β-arrestin1	Purified	N/A
Recombinant bovine β-arrestin2	Purified	N/A
C5aR1pp1	Chemically synthesized	N/A
C5aR1pp2	Chemically synthesized	N/A
C5aR1pp3	Chemically synthesized	N/A
V2Rpp	Chemically synthesized	N/A
CXCR4pp1	Chemically synthesized	N/A
CXCR4pp2	Chemically synthesized	N/A
CXCR4pp3	Chemically synthesized	N/A
CXCR4pp4	Chemically synthesized	N/A
Recombinant human C5a	Purified	N/A
Bradykinin	Genscript	N/A
Arginine Vasopressin Peptide (AVP)	Genscript	N/A
Uranyl formate	Polysciences	Cat# 24762-1
Formvar/carbon coated 300 mesh copper grids	PELCO (Ted Pella)	Cat# 01753-F
Critical commercial assays
Site directed mutagenesis kit	NEB	Cat# E0554
NanoBiT assay	Promega	N/A
Deposited data
C5aR1pp-βarr1-Fab30	This study	PDB: 8GO8, EMD-34173
V2Rpp-βarr2-Fab30	This study	PDB: 8GOC, EMD-34175
C5aR1pp-βarr2-Fab30	This study	PDB: 8GOO, EMD-34178
CXCR4pp-βarr1-Fab30	This study	PDB: 8GP3, EMD-34188
C5aR1pp-βarr1-Fab30-Local-refine	This study	PDB: 8I0N, EMD-35104
V2Rpp-βarr2-Fab30-Local-refine	This study	PDB: 8I10, EMD-35115
C5aR1pp-βarr2-Fab30-Local-refine	This study	PDB: 8I0Z, EMD-35114
CXCR4pp-βarr1-Fab30-Local-refine	This study	PDB: 8I0Q, EMD-35106
Experimental models: Cell lines
Human: HEK293	ATCC	Cat# CRL-3216
Oligonucleotides
C5aR1_R-insertion SDM primer Forward: CGCTCCACAGTGGACACTATGG	This study	N/A
C5aR1_R-insertion SDM primer Reverse: GCGCGTGAATGACTTGCT	This study	N/A
B2Rmut Forward: AACACGGACCTCCATCTCCGTG	This study	N/A
B2RmutReverse GTCATGGAGTTCTCCATCTGAATGGG	This study	N/A
M2Rmut_Forward: AAGTCTACTTCACTGGGCCAC	This study	N/A
M2Rmut_Reverse: GACGGTGTTTTCGTCCTG	This study	N/A
β-arrestin2 Double mutant (βarr2DM) Fw: ggcgAACAACCGTGAAAAACGTG	This study	N/A
β-arrestin2 Double mutant (βarr2DM) Rv: aggaaCGGGGTGATGGTGTAAAC	This study	N/A
Recombinant DNA
PcDNA_3.1 (empty vector)	Dr. Arun K Shukla	N/A
pcDNA3.1_V2R-WT	Dr. Arun K Shukla	N/A
pcDNA3.1_C5aR1-WT	Dr. Arun K Shukla	N/A
pcDNA3.1_CXCR3-WT	Dr. Arun K Shukla	N/A
pcDNA3.1_CXCR7-WT	Dr. Arun K Shukla	N/A
pcDNA3.1_B2R-WT	Dr. Arun K Shukla	N/A
pcDNA3.1_M2R-WT	Dr. Arun K Shukla	N/A
pcDNA3.1_C5aR1-mutTRRST	This study	N/A
pcDNA3.1_B2R-mutΔG368/L370T	Dr. Arun K Shukla	N/A
pcDNA3.1_M2R-mutTVKST	This study	N/A
pCAGGS_V2R-WT-SmBiT	Dr. Arun K Shukla	N/A
pCAGGS _C5aR1-SmBiT	Dr. Arun K Shukla	N/A
pCAGGS_SmBiT-βarr2WT	Dr. Asuka Inoue	N/A
pCAGGS_SmBiT-βarr2DM	This study	N/A
pCAGGS_LgBiT-βarr2WT	Dr. Asuka Inoue	N/A
pCAGGS_LgBiT-βarr2DM	This study	N/A
pCAGGS_LgBiT-FYVE	Dr. Asuka Inoue	N/A
pCAGGS_LgBiT-Ib30	Dr. Asuka Inoue	N/A
Software and algorithms
UCSF Chimera	Pettersen et al.64	https://www.cgl.ucsf.edu/chimera/
UCSF Chimera X	Pettersen et al.65	https://www.rbvi.ucsf.edu/chimerax/
COOT	Emsley et al.66	https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
cryoSPARC	Punjani et al.67	https://cryosparc.com/
PDBsum	Laskowski et al.68	http://www.ebi.ac.uk/thornton-srv/databases/pdbsum/
Phenix	Liebschner et al.69	https://www.phenix-online.org/
PyMol	Schrodinger	https://pymol.org/2/
Prism 8	GraphPad Software	https://www.graphpad.com/scientific-software/prism/
Relion3.1.2	Zivanov et al.70	https://www3.mrc-lmb.cam.ac.uk/relion/index.php?title=Main_Page
SerialEM	Mastronarde71	https://bio3d.colorado.edu/SerialEM/
Graphpad Prism 9	GraphPad Software, San Diego, California USA	https://www.graphpad.com/scientific-software/prism/
ImageJ	Schneider et al.72	https://imagej.nih.gov/ij/download.html