Difference between revisions of "Part:BBa M0050"
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<partinfo>BBa_M0050 short</partinfo> | <partinfo>BBa_M0050 short</partinfo> | ||
| − | This sequence codes for the amino acid sequence AANDENYALAA, which when fused to the C-terminal of proteins, will make the protein susceptible to very fast degradation through SspB-mediated binding to the ClpX protease. | + | This sequence codes for the amino acid sequence AANDENYALAA, which when fused to the C-terminal of proteins, will make the protein susceptible to very fast degradation through SspB-mediated binding to the ClpX protease as well as ClpA protease. |
| − | The following rates of degradation of this tag | + | |
| − | ~5 Vmax/ [Clpx6] min-1 | + | Note that tagged protein degradation rates are dependent on concentration of proteases (ClpX, ClpA) and binding mediators (SspB). Degradation rates are also heavily dependent on protein stability and depend on Km of binding to the protease, which is highly variable with each substrate. Be aware of ClpX, ClpA, and SspB knockouts when using SsrA tags; some expression strains have these mutations. |
| − | ~0.5%/min on log scale from ( | + | |
| − | ~1 min half life from ( | + | The following rates of degradation of respective tagged proteins were observed when this tag was linked to the C-terminal of the protein: |
| + | ~5 Vmax/ [Clpx6] min-1 tagged to Arc from (1), | ||
| + | ~0.5%/min on log scale tagged to GFP from (3), | ||
| + | ~1 min half life tagged to Arc from (2). | ||
See the following references for further information on degradation rates and mechanisms of this tag: | See the following references for further information on degradation rates and mechanisms of this tag: | ||
Revision as of 20:54, 10 December 2007
AANDENYALAA. (Very fast) SsrA degradation tag.
This sequence codes for the amino acid sequence AANDENYALAA, which when fused to the C-terminal of proteins, will make the protein susceptible to very fast degradation through SspB-mediated binding to the ClpX protease as well as ClpA protease.
Note that tagged protein degradation rates are dependent on concentration of proteases (ClpX, ClpA) and binding mediators (SspB). Degradation rates are also heavily dependent on protein stability and depend on Km of binding to the protease, which is highly variable with each substrate. Be aware of ClpX, ClpA, and SspB knockouts when using SsrA tags; some expression strains have these mutations.
The following rates of degradation of respective tagged proteins were observed when this tag was linked to the C-terminal of the protein: ~5 Vmax/ [Clpx6] min-1 tagged to Arc from (1), ~0.5%/min on log scale tagged to GFP from (3), ~1 min half life tagged to Arc from (2).
See the following references for further information on degradation rates and mechanisms of this tag: (1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. (2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]