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bis_104_pq_23_ans_fall_07 - facilitate fusion Additional...

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BIS 104 Practice Question #23 23. Consider the following experiment: You make up two batches of liposomes with identical lipid compositions. The only difference is that batch A has v-SNARE molecules incorporated into the liposomal membrane, whereas, batch B has t-SNARE proteins in the liposomal membrane. The liposomal preparations are mixed together in the following combinations, and you then determine whether or not fusion between mixed liposomes has occurred. v-SNARE liposomes + v-SNARE liposomes t-SNARE liposomes + t-SNARE liposomes t-SNARE liposomes + v-SNARE liposomes mixtures of liposimes, both batches lacking any SNARE proteins. a. Which combinations would you expect to fuse ? only v-SNARE + t-SNARE combinations will fuse; homologous SNARES will not combine and intertwine for fusion. b. What other components might have to be added to the mixed liposome preps in order to
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Unformatted text preview: facilitate fusion? Additional SNAREs; fusogenic proteins; Ca ++ ions. c. What additional proteins are involved in vesicle fusions in vivo? Rabs and tethering proteins. d. What techniques would you use to actually visualize or measure whether or not fusion between liposomes had occurred? The two liposome preps could be made to include two different fluorochromes in their lumen. If fusion occurred, the fluorochromes would intermix and the resulting new color could be seen by fluorescence microscopy. Alternatively, the fluorchromes could be attached to lipids that make up the liposomal membranes. If liposomes with two different fluorochromes fuse, lateral movement resulting in intermixing of the markers could be seen by fluorescence microscopy. (like the Frye and Edidin experiment)...
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