Part:BBa_K4165003
CoH2 (Cohesin)
Cohesin type 2 enzyme binds to its counterpart DocS (BBa_K3396000) to form the protein pair for the PROTAC system.
Usage and Biology
The DocS module comes from The C. thermocellum scaffolding and it could recognize and bind tightly to complementary Coh2 modules. The Coh2–DocS pair represents the interaction between two complementary families of protein modules that exhibit divergent specificities and affinities, ranging from one of the highest known affinity constants between two proteins to relatively low-affinity interactions.CoH2 (Cohesin) is a cellulolytic enzyme in Clostridium thermocellum [1]. The Cohesin family has great affinity for binding with its complementary counterpart family under the name of dockerins with an essential role in the assembly of cellulosomal enzymes into the multienzyme cellulolytic complex (cellulosome). This interaction happens in two different forms, called the dual binding mode, in a calcium-dependent manner due to the presence of a calcium-binding site in the dockerin protein. We used the DocS-Coh2 binding in our protac system to conjugate E3 ligase trim 21 [BBa_K4165001] with the binding peptide for our targeted protein.
Dry Lab
Optimization
This part is considered as an improved version of that of NUDT 2020 team (BBa_K3396001), this part is optimized for expression in E.coli. After optimization, it gave the same protein as that of NUDT after translation.
Modeling
Coh2 has been modeled tagged by GST and His to purify it and increase its stability by GST tag.
Coh2-GST
Figure 1.: Predicted 3D structure of Coh2 protein tagged by GST designed by AlphaFold tool.
Coh2-His
Figure 1.: Predicted 3D structure of Coh2 protein tagged by His designed by TRrosetta.
Dry Lab characterization
PI 4.103 Charge at pH 7 -7.149 Molecular weight 14.741 kDa
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]
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