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Volker Gerke Institute of Medical Biochemistry
ZMBE
Center for Molecular Biology of Inflammation
Membrane Biosynthesis and
Homeostasis
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Biological membranes
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Biological membranes
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Membrane lipids
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Biosynthesis of membrane lipids
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Biosynthesis of glycerophospholipids
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Biosynthesis of glycerophospholipids
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Biosynthesis of glycerophospholipids
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Biosynthesis of glycerophospholipids
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The bilayer transfer problem
Synthesis at cytosolic
leaflet
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Regulated lipid translocation
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Ceramide
Condensation
N-acylation H2O
FA
FA
H2O
Biosynthesis of sphingolipids
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CDP-cholin
Sphingomyelin
UDP-activated sugars
Glycosphingolipids
Ceramide
Biosynthesis of sphingolipids
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Biosynthesis of sphingolipids
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Glycosphingolipids
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Biosynthesis of cholesterol
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Biosynthesis of cholesterol
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Cholesterol incorporation into membranesConsequences: Lowers FA mobility
changes Tm of phase transition
Lowers membrane permeability for small molecules
Supports domain organization
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Asymmetrical distribution of lipids
e.g. plasma membrane
Consequences:
Specific interactions
Signal transduction
Electric contribution to membrane potential
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Biological membranes plasma membrane
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PLCs
PLDs
Inositol and phosphatidylinositol metabolism
Generation of novel membrane binding sites
Generation of phosphatidic acid
Change in membrane potential
Generation of novel membrane binding sites
PLAs Generation of inflammatory mediators
Phospholipid degradation - signal transduction
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Signal transducing lipids
Arachidonic acid
Inflammatory response
Prostaglandine
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Signal transducing lipids - PLC action
PI(4,5)P2
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Signal transducing lipids - PI kinases
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Membrane microdomains (rafts)
Membrane microdomains rich in PIP2, cholesterol, sphingolipids Clustering of these microdomains into larger structures - signalling - membrane transport - lipid driven
- protein driven
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26
Model membranes
Giant unilamellar vesicles (GUV) – 1-100µm lipid composition and curvature chosen to mimick cellular membrane GUV prepared by electroformation between two Indium-tinoxide (ITO) covered slides 1h, T > Tm, lipid , 10Hz, rectangular wave, 1V (peak to peak)
POPC/DOPC/POPS/Chol/PIP2 (38.4:19.2:20:19.4:3)
Intercalating dyes showing a preference for Ld and Lo phases
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DSPC/DOPC/Chol/NBD-DSPE (27:49.9:23:0.1)
GUV
Spontaneous lipid segregation Lo and Ld domains
Visualizing cholesterol – Model membranes
Alexa 594 – Cholesterol analogue
NBD-DSPE
Anna Livia Matos
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PI(4,5)P2
PI(3,4,5)P3
Phosphoinositide segregation in cellular polarity
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Membrane homeostasis Protein/membrane transport in eukaryontic cells
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Principles of vesicle mediated transport
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Vesicle tethering and transport
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Membrane traffic along the secretory pathway
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Membrane traffic along the secretory pathway
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Membrane traffic along the endocytic pathway
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Distribution of phosphoinositides among endocytic compartments
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Domain organization in the endocytic pathway