Difference between revisions of "Part:BBa M0051"

Revision as of 20:58, 10 December 2007

AANDENYNYADAS. (Fast) SsrA degradation tag.

This sequence codes for the amino acid sequence AANDENYNYADAS, a variation of the WT SsrA tag sequence, which, when fused to the C-terminal of proteins, will make the protein susceptible to fast degradation through SspB-mediated binding to the ClpX protease as well as ClpA. DAS variants of SsrA tags require SspB for efficient degradation.

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:~2 Vmax/ [Clpx6] min-1 tagged to Arc from (1).

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_M0051 short</partinfo>
-This sequence codes for the amino acid sequence AANDENYNYADAS, a variation of the WT SsrA tag sequence, which, when fused to the C-terminal of proteins, will make the protein susceptible to  fast degradation through SspB-mediated binding to the ClpX protease. The following rates of degradation of this tag are pulled from the corresponding references below: ~2 Vmax/ [Clpx6] min-1 from (1).
+This sequence codes for the amino acid sequence AANDENYNYADAS, a variation of the WT SsrA tag sequence, which, when fused to the C-terminal of proteins, will make the protein susceptible to  fast degradation through SspB-mediated binding to the ClpX protease as well as ClpA. DAS variants of SsrA tags require SspB for efficient degradation.
+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:~2 Vmax/ [Clpx6] min-1 tagged to Arc from (1).
 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.