KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-DYKDDDDK tag antibody (mouse monoclonal, clone 1E6) | FujiFilm Wako Pure Chemicals | Cat# 012–22384; RRID: AB_10659717 |
| Goat anti-mouse IgG secondary antibody, conjugated with Alexa Fluor 488 | Thermo Fisher Scientific | Cat# A32723; RRID: AB_2633275 |
| Goat anti-mouse IgG secondary antibody, conjugated with Alexa Fluor 647 | Thermo Fisher Scientific | Cat# A21235; RRID: AB_2535804 |
| Anti-β-Arrestin1 antibody (rabbit monoclonal, clone D8O3J) | Cell Signaling Technology | Cat# 12697; RRID: AB_2797996 |
| Anti-β-Arrestin2 antibody (rabbit monoclonal, clone C16D9) | Cell Signaling Technology | Cat# 3857; RRID: AB_2258681 |
| anti-α-tubulin antibody (mouse monoclonal, clone DM1A) | Santa Cruz Biotechnologies | Cat# sc-32293; RRID: AB_628412 |
| Anti-Rabbit IgG, conjugated with horseradish peroxidase | GE Healthcare | Cat# NA9340; RRID: AB_772191 |
| Anti-Mouse IgG, conjugated with horseradish peroxidase | GE Healthcare | Cat# NA9310; RRID: AB_772193 |
| Bacterial and virus strains | ||
| SCS1 Supercompetent Cells | Agilent Technologies | Cat# 200231 |
| E. coli NiCo21(DE3) | NEB | Cat# C2529H |
| E. coli Rosetta (DE3) | Millipore Sigma | Cat# 70954–3 |
| Biological samples | ||
| Biological samples | ||
| Fetal bovine serum | Thermo Fisher Scientific | Cat# 10270–106 |
| Goat serum | Nippon Bio-test Laboratories | Cat# 0208–01 |
| Chemicals, peptides, and recombinant proteins | ||
| Polyethylenimine (PEI) Max (MW 40,000) | Polysciences | Cat# 24765–1 |
| Opti-MEM | Thermo Fisher Scientific | Cat# 31985–070 |
| 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) | Sigma-Aldrich | Cat# H3375 |
| Ethylenediaminetetraacetic acid (EDTA) solution | Thermo Fisher Scientific | Cat# 15575–020 |
| Trypsin | Thermo Fisher Scientific | Cat# 27250–018 |
| Bovine serum albumin | SERVA Electrophoresis | Cat# 11945 |
| Coelenterazine | Carbosynth | Cat# EC14031 |
| Angiotensin II | Peptide Institute | Cat# 4001-v |
| Bradykinin | Peptide Institute | Cat# 4002-v |
| (D-Ala2,N-Me-Phe4,glycinol5)-Enkephalin (DAMGO) | Bachem AG | Cat# 4007829 |
| Dopamine | FUJIFILM Wako Pure Chemical | Cat# 040–15433 |
| Dynorphin A | Peptide Institute | Cat# 4080-v |
| Endothelin-1 | Peptide Institute | Cat# 4198-s |
| Isoproterenol | Tokyo Chemical Industry | Cat# I0260 |
| Isoproterenol | Sigma-Aldrich | Cat# I6504 |
| Methionine-Enkephalin | Peptide Institute | Cat# 4042-v |
| Neurotensin | Peptide Institute | Cat# 4029 |
| Norepinephrine | Tokyo Chemical Industry | Cat# A0906 |
| Oxytocin | Peptide Institute | Cat# 4084-v |
| PAR2 peptide | Peptide Institute | Cat# PAR-3664-PI |
| Parathyroid Hormone (PTH) | Peptide Institute | Cat# 4068-s |
| Serotonin | FUJIFILM Wako Pure Chemical | Cat# 321–42341 |
| Sphingosine 1-phosphate | Cayman Chemical | Cat# 62570 |
| Substance P | Peptide Institute | Cat# 4014-v |
| Thyrotropin-releasing hormone (TRH) | Peptide Institute | Cat# 4011 |
| [Arg8]-vasopressin (AVP) | Peptide Institute | Cat# 4085-v |
| DMEM | Nissui | Cat# 05919 |
| Penicillin G | Sigma-Aldrich | Cat# P3032 |
| Streptomycin | Thermo Fisher Scientific | Cat# 11860–038 |
| ImmunoStar Zeta | FUJIFILM Wako Pure Chemical | Cat# 291–72401 |
| 3-isobutyl-1-methylxanthine (IBMX) | Sigma-Aldrich | Cat# I5879 |
| Forskolin (Fsk) | Sigma-Aldrich | Cat# F6886 |
| V2Rpp (ARGRpTPPpSLGPQDEpSCpTpTApSpSpSLAKDTSS) | Tufts University Core Facility | N/A |
| Monobromobimane (mBBr) | Thermo Fisher Scientific | Cat# M1378 |
| IANBD Amide (N,N’-Dimethyl-N-(Iodoacetyl)-N’-(7-Nitrobenz-2-Oxa-1,3-Diazol-4-yl)Ethylenediamine) | Thermo Fisher Scientific | Cat# D2004 |
| L-Cysteine | Sigma-Aldrich | Cat# 30089 |
| 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB) | Thermo Fisher Scientific | Cat# 22582 |
| BODIPY-TMR PI(4,5)P2 | Echelon Biosciences | Cat # C-45M6 |
| FLAG peptide (DYKDDDDK) | Stanford Pan Facility | N/A |
| AlexaFluor 488 C5 maleimide | Thermo Fisher Scientific | Cat# A10254 |
| ATTO647N C2 maleimide | ATTO TEC | Cat# AD647N-41 |
| Lauryl maltose neopentyl glycol (LMNG) | Anatrace | Cat# NG310 |
| Cholesteryl Hemisuccinate (CHS) | Steraloids | Cat# C6823 |
| Benzamidine hydrochloride hydrate | Sigma-Aldrich | Cat# B6506 |
| Leupeptin | Sigma-Aldrich | Cat# 50-568-49 |
| Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) solution | Sigma-Aldrich | Cat# 646547 |
| ESF921 culture medium | Expression Systems | Cat# 96–001 |
| Ethylenediaminetetraacetic acid (EDTA) | Sigma-Aldrich | Cat# E5134 |
| 2-Mercaptoethanol (BME) | Sigma-Aldrich | Cat# M6250 |
| HBS-P+ Buffer 10x | Cytiva | Cat# BR100671 |
| 08:0 PI(4,5)P2 | Avanti Polar Lipids | Cat# 850185 |
| 08:0 PI(3,4)P2 | Avanti Polar Lipids | Cat# 850183 |
| 08:0 PI(3,5)P2 | Avanti Polar Lipids | Cat# 850184 |
| 08:0 PI(3,4,5)P3 | Avanti Polar Lipids | Cat# 850186 |
| 08:0 PI(3)P | Avanti Polar Lipids | Cat# 850187 |
| 08:0 PI(4)P | Avanti Polar Lipids | Cat# 850182 |
| 08:0 PG | Avanti Polar Lipids | Cat# 840433 |
| Adenosine triphosphate (ATP) | Sigma-Aldrich | Cat# A7699 |
| Isopropyl β-D-1-thiogalactopyranoside (IPTG) | Sigma-Aldrich | Cat# I6758 |
| Neurotensin fragment 8–13 acetate salt | Sigma-Aldrich | Cat# N5266 |
| Series S Sensor Chip SA | Cytiva | Cat# BR100531 |
| Imidazole | Sigma-Aldrich | Cat# I2399 |
| Ampicilin | Sigma-Aldrich | Cat# A9393 |
| Sodium cholate | Sigma-Aldrich | Cat# C6445 |
| HRV3C protease | EMD Millipore | Cat# 71493 |
| Critical commercial assays | ||
| PrimeSTAR HS DNA Polymerase | Takara Bio | Cat# R010B |
| NEBuilder HiFi DNA Assembly Master Mix | New England BioLabs | Cat# E2621X |
| NucleoBond Xtra Midi Plus | Macherey-Nagel | Cat# 740412.50 |
| PureYield Plasmid Miniprep System | Promega | Cat# A1222 |
| FastGene Plasmid Mini Kit | Nippon Genetics | Cat# FG-90502 |
| MycoAlert Mycoplasma Detection Kit | Lonza | Cat# LT07–118 |
| Green Up cADDis cAMP Assay Kit | Montana Molecular | Cat# U0200G |
| BirA500 biotin-protein ligase standard reaction kit | Avidity | Cat #1 |
| Deposited data | ||
| All experimental data | This paper; Mendeley Data | https://dx.doi.org/10.17632/ynrr4j7t4s.1 |
| Active-state crystal structure of β-arrestin 1 bound to the phosphorylated V2R C-tail and Fab30 | Shukla et al., 2013 | PDB: 4JQI |
| Inactive-state crystal structure of β-arrestin 1 | Han et al., 2001 | PDB: 1G4M |
| neurotensin receptor and arrestin2 complex | Huang et al., 2020 | PDB: 6UP7 |
| Experimental models: Cell lines | ||
| Human: HEK293A cells | Thermo Fisher Scientific | Cat# R70507 |
| Human: ΔβArr1/2 DKO HEK293 cells | O’Hayre et al., 2017 | Clones 1 and 2 |
| Insect: Spodoptera frugiperda Sf9 cells | Expression Systems | Cat# 94–001 |
| Insect: Trichuplusia ni Hi5 cells | Expression Systems | Cat# 94–002 |
| Recombinant DNA | ||
| pFastBac-NTSR1 | Huang et al., 2020 | N/A |
| pVL-GRK5 | Huang et al., 2020 | N/A |
| pFB Dual Fab30 | This study | N/A |
| pET15b-β-arrestin-1 | Huang et al., 2020 | N/A |
| pET15b-β-arrestin-1 (ΔCT) | Huang et al., 2020 | N/A |
| pET15b-β-arrestin-1 (3Q) | This study | N/A |
| pET15b-β-arrestin-1 (3Q) (ΔCT) | This study | N/A |
| pET15b-β-arrestin-1 A12C V387C | This study | N/A |
| pET15b-β-arrestin-1 A12C V387C (3Q) | This study | N/A |
| pET15b-β-arrestin-1 L68C | Latorraca et al., 2020 | N/A |
| pET15b-β-arrestin-1 L68C (3Q) | This study | N/A |
| pET15b-β-arrestin-1 L167W L293C | Latorraca et al., 2020 | N/A |
| pET15b-β-arrestin-1 L167W L293C (3Q) | This study | N/A |
| Plasmids for NanoBiT experiments | See Table S4 (Xu et al., 2022; Hisano et al., 2019; Huang et al., 2020; Shihoya et al., 2018; Baidya et al., 2020; Inoue et al., 2012; Kato et al., 2019; Kawakami et al., 2022) | N/A |
| β-arrestin-2-mApple | Eichel et al., 2016 | N/A |
| β-arrestin-2(3Q)-mApple | This study | N/A |
| mApple-N1 | Shaner et al., 2008 | Addgene #54567 |
| Software and algorithms | ||
| GraphPad Prism, Version 8.0, 9.0 | GraphPad, Inc. | https://www.graphpad.com/ |
| Soft Max Pro, Version 7 | Molecular Devices | https://www.moleculardevices.com/products/microplatereaders#softmaxproLink |
| FlowJo software, Version 10 | FlowJo | https://www.flowjo.com/solutions/flowjo |
| PyMOL | Schrö dinger, Inc. | https://pymol.org/2/ |
| FluorEssence Version 3.8 | Horiba, Inc. | https://www.horiba.com |
| Fiji (ImageJ2) Version 2.3.0 | N/A | https://imagej.net/software/fiji/ |
| R studio Version 1.3.1093 | N/A | https://www.rstudio.com/products/rstudio/ |
| R Version 4.0.3 | The R Foundation for Statistical Computing | |
| drc package (in R) Version 3.0–1 | Ritz et al., 2015 | N/A |
| Biacore Control and Analysis Software Version 2.0.4 | Cytiva | N/A |
| MagicPlot Version 3.0 | MagicPlot Systems LLC | https://magicplot.com/ |
| Other | ||
| 96 well white plates | Greiner Bio-one | Cat# 655083 |
| 384 well black | Corning | Cat# 3575 |
| 96 well ½ area white | PerkinElmer | Cat# 6005560 |
| 96-well V-bottom plate | Greiner Bio-one | Cat# 651201 |
| 96-well plate, black | Corning | Cat# 3340 |
| Luminescence microplate reader | Molecular Devices | SpectraMax L, 2PMT |
| Flow cytometer | Sony | EC800 |
| Chemiluminescent imager | Cytiva | Amersham Imager 680 |
| BioTek Multi plate reader | BioTek | BioTek Synergy H4 |
| Tecan multimodal microplate reader | Tecan | Infinite M1000 PRO |
| SPR | Cytiva | Biacore T100 |