Endocytosis and Lysosome Formation The Endolytic Pathway.
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Transcript of Endocytosis and Lysosome Formation The Endolytic Pathway.
Lysosomes and Endosomes
• Early endosome– pH 6.5-6– sorting endosome
• Late endosome– pH 5.5-5
• Lysosome– pH 5-4.5
Structure of Lysosomes• Protein rich membranes
– Lamp/Limp family of glycoproteins
• Unusual lipids– lyso-bisphosphatidic acid
– thought to protect lysosomal membrane lipids from action of lumenal lipases
• Multivesicular bodies (late endosomes/lysosomes)– invagination of the limiting
membrane
– forms internal membranes• TM proteins can segregate into
limiting or internal membranes• limiting membrane can be
recycled, internal is not
Endocytic Mechanisms
• cytockeletal element involvement– macropinocytosis– phagocytosis
• clathrin coated pits• calveolin coated calveolae• non-coated pit pinocytosis
Receptor-Mediated Endocytosis
• Clathrin-coated pits• AP1, 2, 3, 4
– heterotetramer & γ subunits target subunits bind clathrin binds receptor signal sequence
• clathrin triskelion– hexamer of 3 heavy & 3 light chains
Formation of Coated Pit
• AP2-receptor interaction
• Receptor clustering• Clathrin recruitment• Dynamin collar
assembly• Budding• Uncoating & recycling of components
Coated Pit Targeting Sequences
• FxNPxY
• Yxx• Tyrosine residue in these contexts is
necessary & sufficient to mediate AP2 interactions and subsequent clathrin assembly
Endocytosis Kinetics
• Bulk rate– measurement of lipid
movement
• Binding-studies & Turnover– experiments designed to
measure ligand-receptor interaction strength and rate of receptor/ligand destruction, synthesis and replacement
LDL
EGFR
CD4
Pumps
CalveolaeOr as they're better known…
• Plasmalemmal vesicles• Associated with lipid rafts• Membrane lipid cycling• Raft-associated protein
endocytosis/exocytosis– Inner leaflet proteins
• acylated proteins
• prenylated proteins
– Outer leaflet proteins• GPI-linked
– Some TM proteins that interact specifically with glycosphingolipids/cholesterol
Pretty Colored Vesicles
• Gold dust through the endolytic pathway– coated pits (CP)– early endosome (EE)– late endosome (LE)– lysosome (LY)
Recycling Membrane Components from Endosomes
• Tubulovesicular morphology– membrane
concentrated tubules
– bud & recycled directly & indirectly
Ok, It's In the Cell. Now What
• pH dependent interactions– affects protein-protein
interactions in endolytic compartments
• Acidification mediated by H+ ATPase– vacuolar ATPase
Some Examples of Receptor Mediated
Endocytosis• Receptor recycling
– LDL-LDLR– transferrin-transferrin
receptor
• Receptor destruction– EGF-EGFR