CMB Flashcards
GPCR SIGNALLING
- Ligand –> receptor –> conformational change
- GDP exchanged for GTP in α unit –> α unit dissociate from βγ unit
- GTP α unit target protein –> relay signal to other components of signal cascade
- α unit hydrolyse GTP –> GDP = inactivate subunit – this is accelerated by RGS
- Inactive α unit reforms with βγ complex – turn off downstream events
GPCR signalling inactivation
Arrestin – inactivates receptor by preventing receptor interactions with G proteins.
- Prolonged stimulation –> receptors inactivate – receptor kinase (GRK) phosphorylates receptor –> when phosphorylated arrestin binds – inactivates receptor, no interaction with G proteins.
- Receptors into endosomes and be degraded by lysosomes
Receptor tyrosine kinase function
Control cell proliferation, differentiation, survival, metabolism, migration
Receptor tyrosine kinase signalling
- Ligand bind to receptor –> dimerization
- Intracellular tyrosine kinase domains activated –> phosphorylate each other
- Phosphorylated tails recruit adapter protein – Grb-2 & Ras GEF
- Activate Ras protein at the plasma membrane –> ser/thr cascade of phosphorylation
- Phosphorylate MAPKKK –> MAPKK –> MAPK –> phosphorylate bunch of effector proteins
- Proliferation/cell fate determination
Serine threonine kinase signalling
BMP
- Ligand bind to receptor in cell membrane –> receptor subunit 2 to phosphorylate unit 1
- Subunit 1 phosphorylate intracellular smad protein –> bind to another smad protein –> transcriptional regulatory complex
- Enters nucleus –> activate or represses target genes
Absence of Hedgehog signalling
- Membrane protein Patched inhibits the membrane protein Smoothened
- Inhibition of smoothened –> transcription factor Ci being held in cytoplasm
- Ci held in 2 protein complexes. 1 with smoothened, and other with protein suppressor of fused (Su(fu)) - Ci in smoothened complex – phosphorylated by 3 protein kinases – PKA, GSK-3, CK1
- Phosphorylation –> cleavage of Ci –> truncated CiRep
- CiRep –> nucleus and repress Hedgehog target gene
Prescence of Hedgehog signalling
- Hedgehogs bind to patched membrane protein – remove inhibition and block CiRep prod.
- Smoothened is phosphorylated by PKA and CK1
- Ci is released from both complexes in cytoplasm –> enter nucleus – act as gene activator
- Genes activated: wingless, decapentaplegic, and engrailed
Canonical Wnt Signalling
- Wnt absent, β-catenin bound by GSK-3β, CK1-γ (Destruction complex)
- protein kinase phosphorylates β-catenin – target for ubiquitination & degradation in proteasome = no β-catenin free to move into nucleus
- Absence of β-catenin = transcriptional co-repressors bind to TCF transcription factor
- Prevent expression of certain genes
Presence of Wnt Signalling
- Wnt bind to GPCR Frizzled
- Signal across membrane by Frizzled and LRP, activating them
- Activation of Frizzled and LRP cause kinases, GSK-3β, CK1-γ to associate with membrane
- Protein kinases phosphorylate the tail of the activated LRP
- Intracellular signalling protein Dishevelled and protein Axin are recruited to the cytoplasmic tails of LRP and Frizzled.
- Prevent formation of destruction complex –> β-catenin accumulate in the cytoplasm
- β-catenin –> nucleus and bind to TCF, displacing co-repressors –> genes expressed
Intracellular signalling protein Dishevelled in PCP pathway
Dishevelled –> Rho GTPases in cytoplasm – activate proteins that modulate cytoskeletal proteins – affect actin myosin cytoskeleton – affect shape/polarity of cell
Intracellular signalling protein Dishevelled in Ca2+ pathway
Dishevelled –> control molecules willing to release Ca2+ in the cytoplasm – pathway modulating activity of lots of proteins that combine to Ca2+ depends on Ca2+ for activity.
Wnt antagonists
Sfrps
Dkk1
Wif1
Sfrps
structure like frizzled - bind to Wnt - sequester it away from frizzled. Can bind to frizzled and interrupt wnt binding.
Dkk1
binds to LRP, sequester it away from frizzled and Wnt – cannot recruit LRP – pathway not activated
Wif1
Bind to Wnt and sequester them