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| fluoresence from which fluorophore is actually being measured? |
NBD (the green is being measured) |
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mcs5109 Tue, 09 Feb 2010 06:02:39 GMT |
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| Which fluorophore does the quenching and which is quenched? |
-NBD is quenched |
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mcs5109 Tue, 09 Feb 2010 06:02:39 GMT |
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| two fluorophores used to do FRET |
NBD (green)
rhodamine (yellow) |
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mcs5109 Tue, 09 Feb 2010 06:02:39 GMT |
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| FRET |
-florescence resonance energy transfer
- |
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mcs5109 Tue, 09 Feb 2010 06:02:39 GMT |
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| SNAP receptor |
SNARE
-required for fusion
-3 proteins total: 1 in vesicle, 2 in target |
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mcs5109 Tue, 09 Feb 2010 05:58:02 GMT |
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| SNARE docking |
-tight/stable complex
-4 coiled-coils helix bundles
-where E for fusion comes from |
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mcs5109 Tue, 09 Feb 2010 05:58:02 GMT |
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The role of SNAP and NSF is thought to involve dissociation of this complex after
fusion- where does the energy the use to dissociate the complex come from? |
ATP |
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mcs5109 Tue, 09 Feb 2010 05:58:02 GMT |
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| Name the t-SNAREs that act in synaptic vesicle fusion. |
the t-SNAREs syntaxin and Snap25 |
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mcs5109 Tue, 09 Feb 2010 05:44:50 GMT |
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| t-SNARE protein info |
2 polypeptides form the t-SNARE. |
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mcs5109 Tue, 09 Feb 2010 05:44:50 GMT |
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| Name the v-SNARE that acts in synaptic vesicle fusion. |
synaptobrevin |
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mcs5109 Tue, 09 Feb 2010 05:44:50 GMT |
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| The SNARE hypothesis is... |
...an attempt to explain how specificity of docking and fusion is determined |
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mcs5109 Tue, 09 Feb 2010 05:44:50 GMT |
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The luminal space is equivalent to the
_______________. |
extracellular space |
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mcs5109 Tue, 09 Feb 2010 05:31:20 GMT |
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Describe in detail a cycle of nuclear import of a protein with a classical NLS, including
how Ran plays a role. |
yup |
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mcs5109 Tue, 09 Feb 2010 05:31:20 GMT |
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| If a protein wants to know whether it is in the nucleus what does it need to recognize? |
Ran GTP |
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mcs5109 Tue, 09 Feb 2010 05:31:20 GMT |
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| What is the GAP specific for Ran? |
Rna1p in the cytosol |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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| What is the GEF specific for Ran? Where is it? What does it bind? Why is this important? |
Rcc1 in the nucleus |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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| Why is it typically GTP that is bound after the GEF interacts with Ran? |
REPLACES with GTP |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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Understand that the transition from the GTP-bound to GDP-bound state happens
as the result of the enzymatic activity of the G protein (stimulated by the GAP)
In contrast, understand that the transition from the GDP-bound to the GTPbound
state happens as the result of exchange in binding one molecule for another. |
yup |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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| What is a GAP? Do they "turn on" or "turn off" G proteins? |
GTPase activating
protein; turns off |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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| What is a GEF? Do they "turn on" or "turn off" G proteins? |
Guanine nucleotide
exchange factor; turns on |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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| Is the GDP or GTP bound form the active state? |
GTP |
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mcs5109 Tue, 09 Feb 2010 05:27:36 GMT |
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| RanGTP is present in the nucleus due to action of ____________. |
GEF called RCC1 |
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mcs5109 Tue, 09 Feb 2010 05:15:23 GMT |
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which component of the dimeric receptor (alpha/beta) binds the nucleoporin protein in the nuclear
pore? |
beta |
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mcs5109 Tue, 09 Feb 2010 05:15:23 GMT |
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| which component of the dimeric (alpha/beta) receptor binds the NLS? |
alpha |
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mcs5109 Tue, 09 Feb 2010 05:15:23 GMT |
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| transfer of a NLS to a gold bead is an experiment that can demonstrate... |
... that a NLS is sufficient for nuclear localization |
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mcs5109 Tue, 09 Feb 2010 03:46:51 GMT |
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| mutation of a NLS can demonstrate... |
...that the NLS is necessary for nuclear localization |
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mcs5109 Tue, 09 Feb 2010 03:46:51 GMT |
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| what does a classical NLS look like? |
-4 to 8aas
-rich in positively charged aas
-can be situated anywhere in
the protein |
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mcs5109 Tue, 09 Feb 2010 03:46:51 GMT |
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| how big are proteins to cross the nuclear membrane? |
<60kDa |
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mcs5109 Tue, 09 Feb 2010 03:37:18 GMT |
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| What proteins are capable of crossing the nuclear membranes? |
only proteins with the appropriate nuclear localization signal or a small protein, less than 60kD can freely diffuse |
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mcs5109 Tue, 09 Feb 2010 03:40:45 GMT |
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