Virus:Article Title: Ligand recognition and allosteric regulation of DRD1-Gs signaling complexes.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains E. coli cells DH10Bac Invitrogen 10361012 Chemicals, peptides, and recombinant proteins Benzamidine Sigma-Aldrich Cat# B6506 Leupeptin Sigma-Aldrich Cat# L2884 Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace Cat# NG310 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 Cholesterol hemisucinate TRIS Salt (CHS) Sigma-Aldrich Cat# C6512 Sodium Cholate Sigma-Aldrich Cat# C1254 TCEP Thermo Fisher Scientific Cat# 77720 Apyrase NEB Cat# M0398S Ethylenediamine Tetraacetic Acid (EDTA) Thermo Fisher Scientific Cat# S311-500 Monoclonal ANTI-FLAG M2-FITC antibody Sigma- Aldrich Cat# F4049 Fenoldopam Med Chem Express Cat# HY-B0735A A77636 TOCRIS Cat# 1701 Dopamine Selleck Cat# S2529 SKF83959 TOCRIS Cat# 2074 PW0464 This paper N/A LY3154207 This paper N/A [3H]-SCH23390 PerkinElmer NET930250UC Critical commercial assays HTRF Dynamic cAMP kit Cisbio Cat# 62AM4PEB Q5 site directed mutagenesis kit NEB Cat# E0554S cAMP Glosensor Assay Promega Cat# E1291 Deposited data Fenoldopam-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30392 PDB: 7CKW A77636-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30393 PDB: 7CKX PW0464-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30394 PDB: 7CKY Dopamine-PAM-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30395 PDB: 7CKZ SKF83959-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30452 PDB: 7CRH Experimental models: cell lines Insect cell line Sf9 Expression Systems N/A HEK293 cell ATCC N/A Primers for site-direct mutagenesis DRD1-S198A_forward: catctcaGCCtctgtaataagcttttacatccctg This paper N/A DRD1-S198A_reverse: attacagaGGCtgagatggcatatgtcctgctga This paper N/A DRD1-S202A_forward: CtgtaataGCCttttacatccctgtggccatcatg This paper N/A DRD1-S202A_reverse: tgtaaaaGGCtattacagaggatgagatggcatat This paper N/A DRD1-D103A_forward: cctttGCCatcatgtgctccactgcatccatc This paper N/A DRD1-D103A_reverse: agcacatgatGGCaaaggccacccagatg ttacagaag This paper N/A Recombinant DNA pFastBac1-HA-FLAG-DRD1-His*10 This paper N/A pcDNA3.1-HA-FLAG-DRD1 (WT and mutants) This paper N/A (Continued on next page) Cell 184, 943–956.e1–e6, February 18, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER pFastBac1-Gas This paper N/A pFastBac1Dual-Gbg This paper N/A Software and algorithms SerialEM Mastronarde DN https://bio3d.colorado.edu/SerialEM/ MotionCor2 (Zheng et al., 2017) https://emcore.ucsf.edu/ucsf-software Gctf Zhang, 2016 https://www.mrc-lmb.cam.ac.uk/ kzhang/Gctf/ Gautomatch Zhang, 2016 https://www.mrc-lmb.cam.ac.uk/kzhang/ Gautomatch Relion3.0 (Zivanov et al., 2018) https://www2.mrc-lmb.cam.ac.uk/relion CHIMERA Pettersen et al., 2004 http://www.cgl.ucsf.edu/chimera COOT (Emsley and Cowtan, 2004) https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ Phenix Adams et al., 2010 https://www.phenix-online.org PyMOL Schrodinger https://www.pymol.org/2/ Prism v.7.0 GraphPad Software https://www.graphpad.com Other Superdex 200 Increase column 10/300 GE healthcare Cat# 28990944 BIOTEK Cytation3 reader BIO-TEK N/A ESF921 cell culture medium Expression Systems Cat# 96-001 DMEM basic cell culture media GIBCO Cat# 11995 FBS (fetal bovine serum) Cell Box Cat# SAG-01U-02 Penicillin-Streptomycin GIBCO Cat# 15140122 Trypsin 0.5% EDTA GIBCO Cat# 25300054 9 cm2 Cell Culture Dish Corning Cat# 430166 6-well Cell Culture Dish Corning Cat# 430166 96-well Cell Culture Dish Corning Cat# 3599 96-well white microplate assay plates Corning Cat# 3917 ll Article
Recombinant:Article Title: Ligand recognition and allosteric regulation of DRD1-Gs signaling complexes.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains E. coli cells DH10Bac Invitrogen 10361012 Chemicals, peptides, and recombinant proteins Benzamidine Sigma-Aldrich Cat# B6506 Leupeptin Sigma-Aldrich Cat# L2884 Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace Cat# NG310 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 Cholesterol hemisucinate TRIS Salt (CHS) Sigma-Aldrich Cat# C6512 Sodium Cholate Sigma-Aldrich Cat# C1254 TCEP Thermo Fisher Scientific Cat# 77720 Apyrase NEB Cat# M0398S Ethylenediamine Tetraacetic Acid (EDTA) Thermo Fisher Scientific Cat# S311-500 Monoclonal ANTI-FLAG M2-FITC antibody Sigma- Aldrich Cat# F4049 Fenoldopam Med Chem Express Cat# HY-B0735A A77636 TOCRIS Cat# 1701 Dopamine Selleck Cat# S2529 SKF83959 TOCRIS Cat# 2074 PW0464 This paper N/A LY3154207 This paper N/A [3H]-SCH23390 PerkinElmer NET930250UC Critical commercial assays HTRF Dynamic cAMP kit Cisbio Cat# 62AM4PEB Q5 site directed mutagenesis kit NEB Cat# E0554S cAMP Glosensor Assay Promega Cat# E1291 Deposited data Fenoldopam-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30392 PDB: 7CKW A77636-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30393 PDB: 7CKX PW0464-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30394 PDB: 7CKY Dopamine-PAM-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30395 PDB: 7CKZ SKF83959-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30452 PDB: 7CRH Experimental models: cell lines Insect cell line Sf9 Expression Systems N/A HEK293 cell ATCC N/A Primers for site-direct mutagenesis DRD1-S198A_forward: catctcaGCCtctgtaataagcttttacatccctg This paper N/A DRD1-S198A_reverse: attacagaGGCtgagatggcatatgtcctgctga This paper N/A DRD1-S202A_forward: CtgtaataGCCttttacatccctgtggccatcatg This paper N/A DRD1-S202A_reverse: tgtaaaaGGCtattacagaggatgagatggcatat This paper N/A DRD1-D103A_forward: cctttGCCatcatgtgctccactgcatccatc This paper N/A DRD1-D103A_reverse: agcacatgatGGCaaaggccacccagatg ttacagaag This paper N/A Recombinant DNA pFastBac1-HA-FLAG-DRD1-His*10 This paper N/A pcDNA3.1-HA-FLAG-DRD1 (WT and mutants) This paper N/A (Continued on next page) Cell 184, 943–956.e1–e6, February 18, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER pFastBac1-Gas This paper N/A pFastBac1Dual-Gbg This paper N/A Software and algorithms SerialEM Mastronarde DN https://bio3d.colorado.edu/SerialEM/ MotionCor2 (Zheng et al., 2017) https://emcore.ucsf.edu/ucsf-software Gctf Zhang, 2016 https://www.mrc-lmb.cam.ac.uk/ kzhang/Gctf/ Gautomatch Zhang, 2016 https://www.mrc-lmb.cam.ac.uk/kzhang/ Gautomatch Relion3.0 (Zivanov et al., 2018) https://www2.mrc-lmb.cam.ac.uk/relion CHIMERA Pettersen et al., 2004 http://www.cgl.ucsf.edu/chimera COOT (Emsley and Cowtan, 2004) https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ Phenix Adams et al., 2010 https://www.phenix-online.org PyMOL Schrodinger https://www.pymol.org/2/ Prism v.7.0 GraphPad Software https://www.graphpad.com Other Superdex 200 Increase column 10/300 GE healthcare Cat# 28990944 BIOTEK Cytation3 reader BIO-TEK N/A ESF921 cell culture medium Expression Systems Cat# 96-001 DMEM basic cell culture media GIBCO Cat# 11995 FBS (fetal bovine serum) Cell Box Cat# SAG-01U-02 Penicillin-Streptomycin GIBCO Cat# 15140122 Trypsin 0.5% EDTA GIBCO Cat# 25300054 9 cm2 Cell Culture Dish Corning Cat# 430166 6-well Cell Culture Dish Corning Cat# 430166 96-well Cell Culture Dish Corning Cat# 3599 96-well white microplate assay plates Corning Cat# 3917 ll Article
Mutagenesis:Article Title: Ligand recognition and allosteric regulation of DRD1-Gs signaling complexes.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains E. coli cells DH10Bac Invitrogen 10361012 Chemicals, peptides, and recombinant proteins Benzamidine Sigma-Aldrich Cat# B6506 Leupeptin Sigma-Aldrich Cat# L2884 Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace Cat# NG310 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 Cholesterol hemisucinate TRIS Salt (CHS) Sigma-Aldrich Cat# C6512 Sodium Cholate Sigma-Aldrich Cat# C1254 TCEP Thermo Fisher Scientific Cat# 77720 Apyrase NEB Cat# M0398S Ethylenediamine Tetraacetic Acid (EDTA) Thermo Fisher Scientific Cat# S311-500 Monoclonal ANTI-FLAG M2-FITC antibody Sigma- Aldrich Cat# F4049 Fenoldopam Med Chem Express Cat# HY-B0735A A77636 TOCRIS Cat# 1701 Dopamine Selleck Cat# S2529 SKF83959 TOCRIS Cat# 2074 PW0464 This paper N/A LY3154207 This paper N/A [3H]-SCH23390 PerkinElmer NET930250UC Critical commercial assays HTRF Dynamic cAMP kit Cisbio Cat# 62AM4PEB Q5 site directed mutagenesis kit NEB Cat# E0554S cAMP Glosensor Assay Promega Cat# E1291 Deposited data Fenoldopam-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30392 PDB: 7CKW A77636-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30393 PDB: 7CKX PW0464-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30394 PDB: 7CKY Dopamine-PAM-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30395 PDB: 7CKZ SKF83959-DRD1-Gs EM map and coordinate This paper EMDB: EMD-30452 PDB: 7CRH Experimental models: cell lines Insect cell line Sf9 Expression Systems N/A HEK293 cell ATCC N/A Primers for site-direct mutagenesis DRD1-S198A_forward: catctcaGCCtctgtaataagcttttacatccctg This paper N/A DRD1-S198A_reverse: attacagaGGCtgagatggcatatgtcctgctga This paper N/A DRD1-S202A_forward: CtgtaataGCCttttacatccctgtggccatcatg This paper N/A DRD1-S202A_reverse: tgtaaaaGGCtattacagaggatgagatggcatat This paper N/A DRD1-D103A_forward: cctttGCCatcatgtgctccactgcatccatc This paper N/A DRD1-D103A_reverse: agcacatgatGGCaaaggccacccagatg ttacagaag This paper N/A Recombinant DNA pFastBac1-HA-FLAG-DRD1-His*10 This paper N/A pcDNA3.1-HA-FLAG-DRD1 (WT and mutants) This paper N/A (Continued on next page) Cell 184, 943–956.e1–e6, February 18, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER pFastBac1-Gas This paper N/A pFastBac1Dual-Gbg This paper N/A Software and algorithms SerialEM Mastronarde DN https://bio3d.colorado.edu/SerialEM/ MotionCor2 (Zheng et al., 2017) https://emcore.ucsf.edu/ucsf-software Gctf Zhang, 2016 https://www.mrc-lmb.cam.ac.uk/ kzhang/Gctf/ Gautomatch Zhang, 2016 https://www.mrc-lmb.cam.ac.uk/kzhang/ Gautomatch Relion3.0 (Zivanov et al., 2018) https://www2.mrc-lmb.cam.ac.uk/relion CHIMERA Pettersen et al., 2004 http://www.cgl.ucsf.edu/chimera COOT (Emsley and Cowtan, 2004) https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ Phenix Adams et al., 2010 https://www.phenix-online.org PyMOL Schrodinger https://www.pymol.org/2/ Prism v.7.0 GraphPad Software https://www.graphpad.com Other Superdex 200 Increase column 10/300 GE healthcare Cat# 28990944 BIOTEK Cytation3 reader BIO-TEK N/A ESF921 cell culture medium Expression Systems Cat# 96-001 DMEM basic cell culture media GIBCO Cat# 11995 FBS (fetal bovine serum) Cell Box Cat# SAG-01U-02 Penicillin-Streptomycin GIBCO Cat# 15140122 Trypsin 0.5% EDTA GIBCO Cat# 25300054 9 cm2 Cell Culture Dish Corning Cat# 430166 6-well Cell Culture Dish Corning Cat# 430166 96-well Cell Culture Dish Corning Cat# 3599 96-well white microplate assay plates Corning Cat# 3917 ll Article
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