Lecture 2--membranes

2 is their diffusibility affected by their sugar

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Unformatted text preview: al. (2000). Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology 2:288. FRAP-Fluorescence Recovery After Photobleaching A technique that can be used to determine the diffusibility of proteins in the membrane FRAP-Fluorescence Recovery After Photobleaching A technique that can be used to determine the diffusibility of proteins in the membrane the ER recovered faster nuclear pore protiens v ery stable cannot use immunofluorescence becaus eyou are trying to s ee the cell protein moveent and for immunofuorescence you need to fix the cells in place need to use fluorescent protein What factors regulate protein diffusion in the endoplasmic reticulum? Is diffusibility affected by folding status? Is diffusibility affected by sugar modifications? at 59 minutes talked about figure 3 Nehls et al. (2000). Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology 2:288. What factors regulate protein diffusion in the endoplasmic reticulum? VSVG-GFP: An integral membrane protein in the ER that is fused to GFP and expressed in COS cells mutation inVSVG protein that make it unable to fold at high temperature at 32 degree folds normally, at 40 cannot fold; does it look like the folding s tatus of the protein is important for its motility? no, anything fishy about experiment? temperature affects 32ºC, but is easily made the VSVG folds properly at fulidity so could havemisfolded at 40degree version faster and it would have just been due to heat. control\: use v ersion of prtotieins that has no protbelms folding: Is diffusibility affected by folding status? 40ºC Nehls et al. (2000). Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology 2:288. Is diffusibility affected by sugar modifications? Tunicamycin is aaffects the motility of protein. topsugardrug drug that prevents has the synthesis glycosilation and bleacihing does not recover as fast. Nehls et al. (2000). Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology 2:288. What factors regulate protein diffusion in the endoplasmic reticulum? The diffusibility of VSVG in the ER is affected by sugars but not by folding. Nehls et al. (2000). Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology 2:288. Plasma membrane summary: 1. Cell membranes are bilayers of amphipathic lipids 2. The lipid composition of membranes varies 3. Membrane fluidity is regulated by temperature 5. Three types of membrane proteins: 1. Integral, 2. Peripheral, 3. Lipid-anchored 4. Membrane proteins function in signaling, adhesion, and transport 5. The membrane is organized into microdomains 6. Techniques for studying membrane proteins: freeze fracture EM, proteolysis/protection assay, FRAP...
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This note was uploaded on 02/23/2014 for the course MCDB 165A taught by Professor Iruela-arispe during the Winter '08 term at UCLA.

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