Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a The cryo-EM density maps of the D1R-Gs, D2R-Gi, D3R-Gi, D4R-Gi, and D5R-Gs complexes. b The models of the D1R-Gs, D2R-Gi, D3R-Gi, D4R-Gi, and D5R-Gs complexes. c The Ligand-binding pockets the D1R-Gs, D2R-Gi, D3R-Gi, D4R-Gi, and D5R-Gs complexes. d The Rotigotine structure in the D1R-Gs, D2R-Gi, D3R-Gi, D4R-Gi, and D5R-Gs complexes. The EM density of Rotigotine in the five structures is shown.

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Cryo-EM Sample Prep, Ligand Binding Assay

Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a Superposition of the cryo-EM D1R-Gs structures (pink) and the x-ay D1R-G s structure (purple). b Superposition of D1R and D5R structure. c-e Structural features of active Rotigotine-D2R compared with that of bromocriptine-D2R ( a ). Comparison of ligand recognition between Rotigotine-D2R and bromocriptine-D2R ( b ). The structural difference of two active D2R in TM6 and TM7 ( c ). f-h Structural features of active Rotigotine-D3R compared with that of PD128907-D3R ( d ). Comparison of ligand recognition between Rotigotine-D3R and PD128907-D3R ( e ). The structural difference of two active D3R in TM6 and TM7 ( f ).

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Cryo-EM Sample Prep

Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a Detailed interaction between Rotigotine and D1R (left). Effects of mutations of the ligand-binding pocket residues of D1R on changes in ΔpEC 50 in response to stimulation of Rotigotine, evaluated using a GloSensor cAMP assay (right). b Detailed interaction between Rotigotine and D2R (left). Effects of mutations of the ligand-binding pocket residues of D2R on changes in ΔpEC 50 in response to stimulation of Rotigotine, evaluated using a GloSensor cAMP assay (right). c Detailed interaction between Rotigotine and D3R (left). Effects of mutations of the ligand-binding pocket residues of D3R on changes in ΔpEC 50 in response to stimulation of Rotigotine, evaluated using a GloSensor cAMP assay (right). d Detailed interaction between Rotigotine and D4R (left). Effects of mutations of the ligand-binding pocket residues of D4R on changes in ΔpEC 50 in response to stimulation of Rotigotine, evaluated using a GloSensor cAMP assay (right). e Detailed interaction between Rotigotine and D5R (left). Effects of mutations of the ligand-binding pocket residues of D5R on changes in ΔpEC 50 in response to stimulation of Rotigotine, evaluated using a GloSensor cAMP assay (right). All data are presented as mean values ± SEM of three independent experiments for the wild type (WT) and mutants (n=3).

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Ligand Binding Assay, cAMP Assay

Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a Rotigotine competition binding curve to WT Dopamine Receptors. Graphical representation of Rotigotine competition curves for wild-type receptors. Dose response curves were analyzed using “One site – Fit LogIC50” function in Graphpad Prism 8.4 software (Graphpad Software Inc., San Diego, CA). All data are presented as mean values ± standard error of measurement (SEM) with a minimum of four technical replicates and N = 3 biological replicates. b Superposition of Rotigotine structure from the five dopamine receptors. c The binding mode of Rotigotine in D1R. d The binding mode of dopamine in D1R. e Structural comparison of Rotigotine and dopamine in D1R.

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Binding Assay, Software

Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a The chemical structure of Rotigotine. b The interaction of D1R OBP with Rotigotine. c The interaction of D1R EBP with Rotigotine. d Rotigotine affinities (Ki) from radioligand competition binding assays and the OBP-EBP residues alignment. Receptors are listed in order of decreasing Rotigotine affinity. e Structural superposition of five dopamine receptors and bound Rotigotine. f Structural superposition of five dopamine receptors, bound Rotigotine in compared with serotonin receptors (gray): 5-HT 1A (7E2Y), 5-HT 1B (6G79), 5-HT 1D (7E32), 5-HT 2B (6DRY), and 5-HT5A (7X5H). g Structural superposition of five dopamine receptors, bound Rotigotine in compared with adrenergic receptors (gray): a2A (6KUY), a2B (6K41), and a2C (6KUW).

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a Structural superposition of D1R-G s and D5R-G s complex when receptors were aligned. b Comparison of Rotigotine binding poses of D1R and D5R structures. c Structural comparison of Rotigotine recognition between D1R and D5R. d-e Rotigotine-binding pockets of D1R ( d ) and D5R ( e ) viewed from the extracellular side. f Comparison of TM6 and TM7 residues for Rotigotine recognition between D1R and D5R. g The sidechain of W 3.52 residue shows alternative conformation in the D1R-Gs-Rotigotine structure. h The unique conformation of W 3.52 residue is stabilized by a cholesterol molecule in the D5R-Gs-Rotigotine structure. i Comparison of TM3, TM4 and ICL2 residues for compound LY3154207 (a high selective PAM for D1R) recognition between D1R and D5R.

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Structural Genomics of the Human Dopamine Receptor System

doi: 10.1101/2022.10.09.511478

Figure Lengend Snippet: a Cholesterol molecules at the surface of D4R. b A cholesterol molecule is located between TM1 and TM7 of D4R and stabilizes Rotigotine binding through residues W387 7.40 and F91 2.61 . c-f The comparison of the TM1-TM7 structure and residue 2.61 from D1R ( c ), D2R ( d ), D3R ( e ), and D5R ( f ) show difference from D4R.

Article Snippet: Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome.

Techniques: Binding Assay

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Cytotoxicity of Rotigotine in HUVECs. HUVECs were stimulated with 0.1, 0.2, 1, 2, 10, 20, and 100 μM Rotigotine for 24 hours. (a). Molecular structure of Rotigotine; (b). Cell viability (†, ††, †††, P < 0.01, 0.001, 0.0001 vs. vehicle group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques:

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Treatment with Rotigotine reduced ox-LDL-induced expressions of PCSK-9, SREBP-2, and LDL-R in HUVECs. Cells were stimulated with ox-LDL (100 μg/mL) in the inclusion or exclusion of 1 and 2 μM Rotigotine for 24 hours. (a). mRNA of PCSK-9; (b). mRNA of SREBP-2; (c). mRNA of LDL-R; (d). Protein of PCSK-9, SREBP-2, LDL-R (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques:

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Rotigotine decreases ox-LDL-induced total cholesterol and free cholesterol deposition in HUVECs. Cells were stimulated with ox-LDL (100 μg/mL) in the inclusion or exclusion of 1, and 2 μM Rotigotine for 24 hours. (a). Total cholesterol (TC); (b). Free cholesterol (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques:

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Treatment with Rotigotine attenuated ox-LDL-induced oxidative stress in HUVECs. (a). Levels of ROS were measured using DHE staining; (b). m RNA expression of NOX-4; (c). Protein of NOX-4 (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques: Staining, RNA Expression

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Rotigotine inhibited the expression of pro-inflammatory cytokines in HUVECs. (a). TNF-α mRNA; (b). IL-8 mRNA; (c). MCP-1 mRNA; (d). TNF-α secretion as measured with ELISA; (e). IL-8 secretion as measured with ELISA; (f). MCP-1 secretion as measured with ELISA (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Rotigotine suppressed the expression of cell adhesion molecules in HUVECs. (a). VCAM-1 mRNA; (b). ICAM-1 mRNA; (c). VCAM-1 protein as measured with ELISA; (d). ICAM-1 protein as measured by ELISA (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Rotigotine ameliorated ox-LDL- induced attachment of monocytes to HUVECs. Attachment of U937 monocytes to HUVECs was measured using 5-chloromethylfluorescein diacetate (CMFDA) staining (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques: Staining

Journal: Bioengineered

Article Title: Rotigotine protects against oxidized low-density lipoprotein(ox-LDL)-induced damages in human umbilical vein endothelial cells(HUVECs)

doi: 10.1080/21655979.2021.2000224

Figure Lengend Snippet: Rotigotine inhibited activation of NF-κB. (a). Levels of nuclear NF-κB p65; (b). Luciferase activity of NF-κB (†††, P < 0.0001 vs. vehicle group; ǂ, ǂǂ, P < 0.01, 0.001 vs. ox-LDL group, N = 5–6)

Article Snippet: Cells were exposed to ox-LDL (100 μg/ml) (Sigma-Aldrich, USA) in the inclusion or exclusion 1 and 2 μM Rotigotine (Topscience, Shanghai, China) for another 24 hours [ ].

Techniques: Activation Assay, Luciferase, Activity Assay

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Repurposing DrugBank compounds as potential Plasmodium falciparum class 1a aminoacyl tRNA synthetase multi-stage pan-inhibitors with a specific focus on mitomycin

doi: 10.1016/j.ijpddr.2024.100548

Figure Lengend Snippet: Binding energies, selectivity index (SI) and pan SI (PSI) values for the 7 identified DrugBank compounds of class 1a tRNA synthetases.

Article Snippet: Only three ( MMC , bicalutamide, and rotigotine ) of the seven hit compounds were available for purchase (MolPort; https://www.molport.com/shop/index ).

Techniques: Binding Assay

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Repurposing DrugBank compounds as potential Plasmodium falciparum class 1a aminoacyl tRNA synthetase multi-stage pan-inhibitors with a specific focus on mitomycin

doi: 10.1016/j.ijpddr.2024.100548

Figure Lengend Snippet: Summary of the in vitro antiplasmodial activities of the hit compounds identified in silico .

Article Snippet: Only three ( MMC , bicalutamide, and rotigotine ) of the seven hit compounds were available for purchase (MolPort; https://www.molport.com/shop/index ).

Techniques: In Vitro, In Silico, Inhibition, Control