Part:BBa_K5301001
SdyCatcher can achieve covalent binding of proteins through Tag-Catcher interaction.
SdyCatcher is a fibronectin-binding protein. It can cooperate with SdyTag to achieve covalent binding of proteins through Tag-Catcher interaction. SdyCatcher and SdyTag 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 SdyCatcher with spNW15, SpyCatcher and mCherry[1-10] (namely SCSdC-mCh[1-10]). We employed SdyTag and SdyCatcher to link two proteins - SCSdC-mCh[1-10] and SnCSdT.
Characterization
We used SDS-PAGE to test whether the protein containing SdyCatcher had been expressed successfully(Figure 1). The molecular weight of SCSdC-mCh[1-10] (containing SdyCatcher) is 76.3 kDa. And we purified the target protein with a molecular weight of approximately 70-100 kD (Lane 4 - Lane 5), which demonstrated that we had successfully expressed the protein containing SdyCatcher.
Figure 1.SDS-PAGE analysis of the extraction results of SCSdC-mCh[1-10] (containing SdyCatcher).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 SCSdC-mCh[1-10] is 76.3 kDa.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 10
Illegal AgeI site found at 58 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 55
Illegal BsaI.rc site found at 256
- ↑ 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.
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