User:Jaime B. Hutchison/Sandbox 1
From Proteopedia
(Difference between revisions)
| Line 2: | Line 2: | ||
| + | The biologically active form of this protein is proposed to be a | ||
| + | <scene name='Sandbox_Reserved_126/Dimer/1'>homodimer</scene>. | ||
| - | + | We would like to make fluorescent mutants for microscopy applications. One way to do this is by attaching maleimide dye to Cys amino acids present in the protein. The homodimer has | |
| - | + | ||
| - | The | + | |
<scene name='Sandbox_Reserved_126/Dimer/2'>4 cysteine (Cys) amino acids</scene>. | <scene name='Sandbox_Reserved_126/Dimer/2'>4 cysteine (Cys) amino acids</scene>. | ||
(<font color='orange'>'''Cys is orange'''</font>.) | (<font color='orange'>'''Cys is orange'''</font>.) | ||
| - | + | Before designing the mutant we need to check that there are | |
| - | <scene name='Sandbox_Reserved_126/Dimer/4'>Cys sites available on the surface</scene>. | + | <scene name='Sandbox_Reserved_126/Dimer/4'>Cys sites available on the surface</scene> for maleimide attachment. |
| - | Because Cys 82 looks to be accessible and because the placement of Cys 66 looks like attaching a fluorophore to it may interfere with membrane binding we will mutate | + | Because Cys 82 looks to be accessible and also because the placement of Cys 66 looks like attaching a fluorophore to it may interfere with membrane binding we will mutate |
<scene name='User:Jaime_B._Hutchison/Sandbox_1/Dimer/1'>Cys 66</scene>. | <scene name='User:Jaime_B._Hutchison/Sandbox_1/Dimer/1'>Cys 66</scene>. | ||
This will be done for each monomer, | This will be done for each monomer, | ||
| Line 17: | Line 17: | ||
<scene name='User:Jaime_B._Hutchison/Sandbox_1/Dimer/8'>alanines (Ala)</scene>. | <scene name='User:Jaime_B._Hutchison/Sandbox_1/Dimer/8'>alanines (Ala)</scene>. | ||
(<font color='purple'>'''Ala is purple'''</font>.) | (<font color='purple'>'''Ala is purple'''</font>.) | ||
| + | Each of the mutated monomers will now have only one binding site at | ||
| + | <scene name='User:Jaime_B._Hutchison/Sandbox_1/Dimer/9'>Cys 82</scene>. | ||
Revision as of 19:08, 3 June 2010
|
The biologically active form of this protein is proposed to be a
.
We would like to make fluorescent mutants for microscopy applications. One way to do this is by attaching maleimide dye to Cys amino acids present in the protein. The homodimer has . (Cys is orange.) Before designing the mutant we need to check that there are for maleimide attachment. Because Cys 82 looks to be accessible and also because the placement of Cys 66 looks like attaching a fluorophore to it may interfere with membrane binding we will mutate . This will be done for each monomer, . The mutation will replace the cysteines at residue 66 with . (Ala is purple.) Each of the mutated monomers will now have only one binding site at .
