Difference between revisions of "Part:BBa K5301002"
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Revision as of 09:41, 1 October 2024
SdyTag can achieve covalent binding of proteins through Tag-Catcher interaction.
SdyTag is a fibronectin-binding protein. It can cooperate with SdyCatcher to achieve covalent binding of proteins through Tag-Catcher interaction. SdyTag and SdyCatcher can also be used alongside SpyTag-SpyCatcher partners, to demonstrate kinetically controlled directed protein ligation, domain specific protein circularization and macromolecular assembly [1].
Usage and Biology
We devised the fusion expression of SdyTag with spNW15 and SnoopCatcher (namely SnCSdT). We employed SdyTag and SdyCatcher to link two proteins - SCSdC-mCh[1-10] and SnCSdT.
We also employed AlphaFold to predict the structure of the protein constituted by SdyTag, spNW15, and SdyCatcher, and discovered that the SdyTag was successfully bound to SdyCatcher(Figure 1).
Figure 1.The structure of the protein constituted by SdyTag, spNW15 and SdyCatcher as predicted by AlphaFold
Characterization
We used SDS-PAGE to test whether the protein containing SdyTag had been expressed successfully(Figure 2). The molecular weight of SnCSdT (containing SdyTag) is 42.1 kDa. And we purified the target protein with a molecular weight of approximately 43 kD (Lane 5 - Lane 8), which demonstrated that we had successfully expressed the protein containing SdyTag.
Figure 2.SDS-PAGE analysis of the extraction results of SnCSdT (containing SdyTag).The gel was run at 80 V for 10 minutes and then at 150 V for 20 minutes, followed by staining with Coomassie Brilliant Blue. The molecular weight of SnCSdT-M15-GS is 42.1 kDa.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
- ↑ Tan, L.L., S.S. Hoon and F.T. Wong, Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly. PLOS ONE, 2016. 11(10): p. e0165074.