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This document discusses intracellular receptors as drug targets. It focuses on the estrogen receptor. Intracellular receptors contain zinc fingers that allow interactions with zinc and DNA binding. Their mechanism involves a messenger crossing the cell membrane and binding to the receptor. The receptor then dimerizes and binds to co-activator proteins before binding to DNA to regulate transcription. The estrogen receptor binds the messenger estradiol in its binding pocket through important phenol and alcohol groups on estradiol. Antagonists like raloxifene and tamoxifen mimic estradiol's binding but prevent the receptor from adopting a conformation that recruits co-activators and promotes transcription.
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Contents
Part 4: Section 6.8
5. Intracellular receptors
5.1. Structure
5.2. Mechanism
5.3. Oestrogen receptor (5 slides)
[8 slides]](data:image/gif;base64,R0lGODlhAQABAIAAAAAAAP///yH5BAEAAAAALAAAAAABAAEAAAIBRAA7)
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CELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTION
CELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTIONCELL MEMBRANE , RECEPTOR , DRUG RECEPTOR INTERACTION
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1. Protein purification involves isolating one or a few proteins from a complex mixture through a series of extraction, precipitation, ultracentrifugation, and chromatographic steps.
2. Maintaining the proper buffering environment is essential to prevent changes in pH that could denature proteins. Buffers should be selected so their pH is within 1 unit of the protein's pKa.
3. When proteins are purified, they should be stored with additives at the appropriate pH and temperature depending on the protein's stability requirements for downstream applications. Storage conditions need to be optimized for each protein.
Methodology of purification and characterization of receptors
Methodology of purification and characterization of receptorsCentral University of Gujarat, Gandhinagar
This document discusses the methodology for purifying, reconstituting, and characterizing receptors and synthetic analogues of epinephrine. It describes how receptor proteins are isolated from natural sources through cloning or from cell membranes using techniques like ultrasonication and centrifugation. The isolated receptors are then solubilized using detergents and purified using affinity, ion exchange, or size exclusion chromatography. The purified receptors can be reconstituted into detergent micelles, bicelles, or nanodiscs to mimic the cell membrane environment. Characterization of the receptors involves studying their primary structure, post-translational modifications, ligand binding sites, and interactions with G-proteins. Synthetic analogues of epinephrine are also designed to mimic itsSimilar to patrick_ch06_p4.ppt (20)
Affinity chromatography by Shiv kalia ( m.pharma analytical chemistry)
Affinity chromatography by Shiv kalia ( m.pharma analytical chemistry)
cell communications and cellular signalling systems vishnu
cell communications and cellular signalling systems vishnu
Medicine Level 1 Physiology: Cellular Communication
Medicine Level 1 Physiology: Cellular Communication
Methodology of purification and characterization of receptors
Methodology of purification and characterization of receptors
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Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
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2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
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5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
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- 1. 1 © Patrick An Introduction to Medicinal Chemistry 3/e Chapter 6 PROTEINS AS DRUG TARGETS: RECEPTOR STRUCTURE & SIGNAL TRANSDUCTION Part 4: Section 6.8
- 2. 1 © Contents Part 4: Section 6.8 5. Intracellular receptors 5.1. Structure 5.2. Mechanism 5.3. Oestrogen receptor (5 slides) [8 slides]
- 3. 1 © 5. Intracellular receptors • Chemical messengers must cross cell membrane • Chemical messengers must be hydrophobic • Example - steroids and steroid receptors
- 4. 1 © 5.1 Structure Zinc Zinc fingers contain Cys residues (SH) Allow S-Zn interactions CO2H H2N DNA binding region (‘zinc fingers’) Steroid binding region 5. Intracellular receptors
- 5. 1 © Cell membrane 5.2 Mechanism 1. Messenger crosses membrane 2. Binds to receptor 3. Receptor dimerisation 4. Binds co-activator protein 5. Complex binds to DNA 6. Transcription switched on or off 7. Protein synthesis activated or inhibited Messenger Receptor Receptor-ligand complex Dimerisation Co-activator protein DNA 5. Intracellular receptors
- 6. 1 © 5.3 Oestrogen receptor Oestradiol H12 Oestrogen receptor Binding site AF-2 regions Dimerisation & exposure of AF-2 regions Coactivator Nuclear transcription factor Coactivator DNA Transcription 5. Intracellular receptors
- 7. 1 © 5.3 Oestrogen receptor O Me OH H H H H H H2O His 524 Glu353 Arg394 Hydrophic skeleton Oestradiol • Phenol and alcohol of oestradiol are important binding groups • Binding site is spacious and hydrophobic • Phenol group of oestradiol positioned in narrow slot • Orientates rest of molecule • Acts as agonist 5. Intracellular receptors
- 8. 1 © 5.3 Oestrogen receptor OH S O O Raloxifene Asp351 His 524 O Glu353 Arg394 N H H Side chain • Raloxifene is an antagonist (anticancer agent) • Phenol groups mimic phenol and alcohol of oestradiol • Interaction with Asp351 is important for antagonist activity • Side chain prevents receptor helix H12 folding over as lid • AF-2 binding region not revealed • Co-activator cannot bind 5. Intracellular receptors
- 9. 1 © Tamoxifen (Nolvadex) - anticancer agent which targets oestrogen receptor CH2CH3 O Me2N 5.3 Oestrogen receptor 5. Intracellular receptors