3 - G-Proteins + Calcium Flashcards
How can you turn off a G-Protein?
GTPase in alpha subunit. Prevents overreactive. Drugs and toxins can target GTPase
How would you describe a G-protein?
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What is the main set up of a GPCR?
- A receptor (ligand bind in TMD or N terminal)
- G-protein
- Effector (enzyme/ion channel)
- Second messenger
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What are the three types of G-protein and what do they govern?
- Gs - Stimulate adenylate cyclase
- Gi - Inhibit adenylate cyclase
- Gq - Stimulate phospholipase C
What types of G-protein are all the main receptors?
M2 also salivary glands
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What is the cascade pathway of a Gs protein?
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What is the cascade pathway of a Gq protein?
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How does a cholera and pertussis toxin work?
Pertussis - Gi protein. Covalently binds to alpha subunit and inhibits GDP exchange for GTP
Cholera - Prevents GTPase action so Gs-protein constantly activated, can lead to loss of Cl- from cells, leading to diarrheoa
What is the cascade pathway of a Gi protein?
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How does Phospholipase C work?
- Cuts PIP2, (membrane phospholipid), into DAG and IP3.
- IP3: Water soluble so binds to LGIC on ER. Receptor stimulated and opens. Influx of calcium ions into cytosol
- DAG: Lipid soluble. Activates PKC, which acts on lots of proteins
How does cyclic AMP activate PKA?
PKA has 4 subunits. cAMP binds to R subunits and releases C subunits so they are active
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How does positive inotropy in the heart work?
- B1 Adrenoreceptor therefore GS
- Sympathetic adrenaline
- PKA phosphorylates VOCC so that when there is wave of depolarisation, more Ca2+ enters the cell, more Ca2+ released from ER, more forceful contraction
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What G protein is involved in smooth muscle contraction?
Q
- Vasoconstriction - a1
- Bronchoconstriction - M3
- GI Tract - a2?
How does arteriolar vasoconstriction work?
- Sympathetic noradrenaline on a1 receptors
- IP3 produced which releases Ca2+ from ER, leading to contraction
- DAG produced activates PKC which can activate proteins that sustain contraction
How do you modulate neurotransmitter release?
- U-opiod Gi receptor
- BY unit binds to VOCC stopping it from opening
- Less Ca2+ in cell, therefore vesicles containing neurotransmitter cannot be released
Where are U-Opioid receptors found?
In presynaptic neurones
How is chronotropy regulated in the heart?
- M2 so Gi
- Production of BY binds to K+ channels so membrane more permeable to K.
- Slows polarisation of membrane as K is leaving
- SA fires slower, negative chronotropy
If opioids stop VOCC working, how does the heart keep contracting?
VOCC in neurones are a different type to in the heart.
Heart - L
Neurones - P/Q and N
Why is signal amplification important?
Allows small changes in extracellular signals to cause significant changes in intracellular behaviour
What are serum calcium levels?
8-10 mg/dL. 50% is free
How is calcium homeostasis regulated?
- Intestinal calcium uptake
- Bone calcium regulation
- Reabsorption in kidneys
ENDOCRINE CONTROL
What is the concentration of calcium in different compartments?
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What hormones regulate calcium levels?
- Calcitonin (thyroid)
- Parathyroid hormone
- Ca receptors in parathyroid gland
- 1,25 Dihydroxyvitamin D3
Why are calcium gradients important?
- Muscle contraction
- Fertilisation
- Cell apoptosis
- Neurotransmission
How is calcium concentration increased in a cell?
- LGIC
- VOCC
- IP3
- CICR (ryanodine receptor) (calcium binds to receptor opening it)
- SOCC (low Ca in ER, ligand from ER binds to channel)
How are calcium gradients set up in a cell?
- NCX
- PMCA
- SERCA
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Why is NCX classed as active if it is using Na’s concentration gradient?
Na+/K+ ATPase is setting up concentration gradient of sodium actively
What are the two responses in intracellular calcium concentration when a receptor is activated and why are there multiple responses?
- Peak Plateau
- Oscilliations
Some proteins need large amounts of calcium to be acitvated, some small and calcium increase may be within microdomains not across whole cell
What G proteins does the pertussis toxin act on and why doesnt it affect GS?
- Gi and Go
- Doesn’t affect Gs as doesnt have same alpha subunit