GPCr detail

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Last Class: A. intracellular vesicle traffic 1. ER to golgi 2. endocytosis, exocytosis B. Signaling Transduction 1. Ligand Receptor Interactions, 2. Intracellular signaling molecules, 3. molecule switches: phosphorylation and GTP binding, 4. Signaling integration, 5. complex formation, 6. Signaling amplification, 7. Signaling desensitization

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Cell Communication Different Receptor Types Ion channel-linked receptors *GPCR signaling *Enzyme linked receptors

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G Protein Coupled Receptors (GPCR)

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A G-Protein-Coupled Receptor Or G Protein-linked Receptor 7 transmembrane domains

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The disassembly of G-Protein upon stimulation Spontaneous deactivation is very fast, in minutes. However, with the help of RGS (regulator of G protein signaling, a GAP for a unit), signals can be shut off even faster

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The Activation cycle of G-Protein

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GPCR Signaling: cAMP

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The visualization of cAMP in nerve cells GPCR->Gs->adenylyl cyclase->cAMP Gi

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cAMP cycle: GPCR->Gs->adenylyl cyclase->cAMP Cyclic AMP phosphodiesterase breaks down cAMP to 5 ’ -AMP

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The function of cAMP Targeting PKA (cyclic-AMP-dependent protein kinase A)

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The Whole Signaling Network related to cAMP

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Terminology: CRE(cyclic AMP response element); CREB: CRE binding protein; CBP: CREB binding protein

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GPCR Signaling: Calcium

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Three Types of Inositol phospholipids PI, PI(4)P, PI(4,5)P2

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Phospholipase C- b (PLC- b ) Produces DAG (diacylglycerol) and IP3 (inositol 1,4,5-trisphosphate (IP3)) Gq->PLC- b

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Gq signaling pathways and Calcium

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Fertilization of an egg by a sperm triggering an increase in cytosolic Calcium 3 major types of calcium channels: Voltage dependent Ca channels on plasma membrane IP3-gated Ca release channels on ER membrane Ryanodine receptor on ER membrane

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Calcium uptake and deprivation Na/Ca exchanger on plasma membrane, 2. Ca pump on ER membrane, 3. Ca binding molecules, 4. Ca pump on Mitochondia

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Calcium Frequency encoding signaling strength Local Ca blips, sparks, puffs, reflecting local opening of individual channels in ER, strong local signal induces global activity, the elevated Calcium trigger calcium deprivation system

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Targeting molecules for Calcium Calcium binding protein Calmodulin

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Ca 2+ /calmodulin dependent protein kinase (CaM-kinase) Memory function: 1. calmodulin dissociate after 10 sec of low calcium level; 2. remain active after calmodulin dissociation

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Ca 2+ /calmodulin dependent protein kinase (CaM-kinase) Frequency decoder of Calcium oscillation High frequence, CaM-kinase does not return to basal level before the second wave of activation starts

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Desensitization of GPCR 1. Inhibitory structural alteration of receptor; 2. receptor internalization; 3. receptor degration GRK (G protein-linked receptor kinase) Arrestin takes to clathrin-coated pits and degradation

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GPCR Signaling Summary 1. G-protein types 2. cAMP and Calcium signaling pathways 3. desensitization

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Enzyme-Linked Cell Surface Receptors *Receptor Tyrosine Kinase *Tyrosine kinase associated receptors *Receptor-like tyrosine phosphatase *Receptor serine/threonine kinase Receptor guanylyl cyclase Histidine like associated receptor

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Receptor Tyrosine Kinases (RTKs)

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Seven subfamilies of receptor tyrosine kinases

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Three ways in which signaling proteins can cross-link receptor chains 1. dimer, 2. monomer but brought together by proteoglycan, 3. cluster on membrane

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The importance of receptor oligomerization

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The docking of signaling molecules at RTK

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The binding of SH2-containing intracellular signaling proteins to an activated PDGF receptor

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The structural view of SH2 domain

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RTK Signaling: Ras Pathway

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The regulation of Ras activity, a famous downstream molecule of RTK responsible for cancer development

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The activation of Ras by RTK signaling

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The MAP-kinase regulated by Ras

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RTK Signaling: PI3K Pathway

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The inositol phospholipids generated by PI3K

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The recruitment of signaling molecules with PH domains to the plasma membrane during B cell activation One PI3K pathway PH domain: pleckstrin homology domain

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Another PI3K pathway to regulate cell survival

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Another PI3K pathway to regulate cell migration PI3K->PIP3->GEF->Rac->Wave->Arp2/3->Actin polymerization Movie: chemotaxis, PI3K and cell migration

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Intracellular Signaling Pathways activated by RTKs and GPCRs

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Tyrosine kinase associated receptors Integrins: cell-extracellular matrix adhesion Binding to Src and FAK

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Receptorlike tyrosine phosphatases Intracellular protein and receptor

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Receptor Serine/threonine kinase Transforming growth factor (TGF-b) and Smad signaling pathway

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Protein kinase Summary

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Enzyme-linked Receptor Signaling Summary 1. receptor types 2. RTK and its signaling: Ras and PI3K 3. Tyrosine kinase associated receptors and Receptor-like tyrosine phosphatase 4. Receptor serine/threonine kinase, TGF- b and Smad

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Other Signaling Pathways Proteolysis mediated

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Notch and Delta interaction Lateral inhibition

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The inhibitory pathway of Notch Proteolysis-mediated

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The inhibitory pathway of Notch Proteolysis-mediated

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Wnt Signaling pathway LRP: LDL-receptor-related protein; GSK-3 b : glycogen synthase kinase-3 b; APC: adenomatous polyposis coli;

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NF-KB pathway

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Summary 1. GPCR signaling: PKA and Calcium 2. Enzyme-linked Receptor signaling: RTK->Ras and PI3K 3. Proteolysis-mediated signaling pathways