1. endothelial growth factor (EGF)
-tyrosine kinase in EGF-receptor is phosphorylated, then Obl protein binds to tyrosine kinase domain as to target EGF-receptor for degradation; downregulation of EGF-receptor --> fuse into late endosome via inward budding mechanism, resulting multivesicular body or ESCRT pathway, fuse into lysosome .:degradation:.
-cf. to mutated tyrosine kinase region = will be recycled back to plasma membrane instead of being degraded in lysosome
2. rab5, rabex5/rabaptin5 complex, PI3P:
-rab5 assay in EE to EE fusion; avidin-biotin peroxidase
-rab5 recruits rabex5/rabaptin5 complex (feedback loop 1), recruits vsp34 kinase enzyme to catalyze PI to PI3P, to enable EEA1 binds to rab5 (feedback loop2)
-rab effector protein in vitro assay: rab5gtp column, add with cytosol, elute the bound specific protein
3. PIP2 in budding, coating and uncoat
- budding: amphiphysin recruits AP2&dynamin, then dynamin binds to PIP2, then epsin binds to AP2 and PIP2 = invagination/budding
-coating: AP180 binds to PIP2 and clathrin, Ap180 interacts with AP2 = clathrin coated pit
-uncoat: synaptojanin dephosphorylate PIP2 at 5th position, recruitment Hsc70ATPase & auxillin = uncoat
