Lecture 9 - Tyrosine Kinase Signaling Flashcards
What are the 2 types of tyrosine kinases?
- Receptors
2. Receptor-associated
List 5 receptor tyrosine kinases.
- Insulin receptor
- IGF-1 receptor
- Nerve growth factor receptor
- Epidermal growth factor receptor
- Platelet derived growth factor
+ many more growth factor receptors
What must be true of receptor associated tyrosine kinases?
They are soluble in the cytoplasm
What do the receptor tyrosine kinases have in common? 3 parts
- 1 single transmembrane domain
- Elaborate, large, and globular extracellular domain
- Intracellular tyrosine kinase catalytic domain
Where is the catalytic domain of receptor tyrosine kinases located?
Proximal to the membrane on the cytosolic side
Describe the 4 steps of receptor tyrosine kinase activation.
- Ligand binding induces receptor dimerization through various mechanisms
- Dimerization activates the tyrosine kinase domain of the receptor by moving the regulatory binding from the active site
- Tyrosine kinase INTERmolecularly trans-autophosphorylates the receptor mostly OUTSIDE of the catalytic domain
- Substrate phosphorylation
Which ONLY receptor tyrosine kinase is a dimer? How?
What is also unique about this receptor’s functioning?
Insulin receptor is covalently dimerized
Also phosphorylates insulin receptor substrates (IRS) on tyrosines, creating even more phosphotyrosine binding sites, on top of the usual binding sites on the receptor itself
Is the trans-auto-phosphorylation of receptor tyrosine kinase necessary for proper substrate phosphorylation?
YUP
What is the most fundamental difference between tyrosine kinases signaling strategies and Ser/Thr kinases signaling strategies?
- Ser/Thr kinases: presence of P on substrates results in a conformational change which activates or inactivates(couple of exceptions)
- Tyr kinases: some small conformational changes, but mainly the cytosolic phosphotyrosines serve as binding sites for phosphotyrosine binding proteins
What are 3 examples of phosphotyrosine binding proteins?
- PI3-K using a p85 adaptor
- GAP = GTPase Activating Protein
- PLPC-gamma
Describe the specificity of the phosphotyrosine binding sites and give a supporting argument.
Each binding protein has a VERY specific recognition 5 AA sequence including the phosphotyrosine
Synthetic peptides, as short as 5 AAs, containing a P-tyrosine can block binding to SPECIFIC binding sites on the receptor
Why did researchers originally think the tyrosine kinases had low affinity and specificity? How is this overcome by the tyrosine kinase?
Because they have low affinity and specificity at their catalytic site, not their phosphotyrosine binding sites:
- Deal with lack of specificity by having the high specificity at the 5 AA sequences
- Deal with lack of affinity by bringing in the substrates to the enzyme, creating an artificially high substrate concentration to work with the high Km of the catalytic site
What parts of the phosphotyrosine binding proteins bind to the binding sites?
Src homology 2 domains (SH2)
2 types of receptor associated protein tyrosine kinases?
- Src
2. Jak/STAT
To what kind of receptors do receptor associated protein tyrosine kinases bind? 6 examples?
Cytokine receptors
Eg: GH receptor, prolactin receptor, interferon receptor, erythropoietin receptor, G-CSF receptor, IL receptors