Part:BBa_K2923011
MS2 coat protein
MS2 is a coat protein from the Enterobacteria phage MS2. This virus is a RNA virus and this protein can fix a specific RNA stem loop
Improvement by AFCM-Egypt 2024
This MS2 coat protein (MCP) exhibits a strong affinity for a specific stem-loop structure (MS2). This binding ability makes it a valuable tool for studying RNA localization, purification, and interactions. It can be used for RNA Visualization By fusing it to fluorescent protein which in turn can be used to study RNA-protein interactions.
In our model, we have added a new feature to MCP (BBa_K2923011) ,which was designed by iGEM19_Strasbourg to act as a protein sensor that could recruit different effector proteins to its specific stem loop (MS2) .
While it originally relied solely on its high binding affinity to MS2 RNA, we have implemented MCP in our metalloproteinase-9 (MMP-9) based mRNA switch by incorporating a specific sensor (nanobody) that recognizes MMP9, a structural protein that increases significantly within cells during tissue injury like burns. Nanobodies are a unique type of antibody which is used to recognize specific types of proteins making them ideal for applications in diagnostics and therapeutics This modification allows our system to switch from an inactive to an active state in response to tissue damage. Moreover, MCP could act as an intracellular protein sensor by combining MCP to a nanobody specific to the protein of interest.
)this figure illustrates our improvement on MCP by combining it to nanobody to act as an intracellular protein sensor. .
=Characterization by AFCM-Egypt2024
Dry lab Characterization by AFCM-Egypt 2024
We measured the effect of the nanobodies on the MCP-MS2 binding stability, so we measured MCP-MS2 before and after binding to nanobody1
MCP-MS2 binding stability
Alignment Plot
3D structure of MCP binded to MS2-Aptamers
this figure show That the alignment plot scores high diagonal intensity which indicates the similarity between our structures and the experimental one .
MS2-MCP-NB1 binding stability
Alignment Plot
3D structure of MCP binded to MS2-Aptamers and NB1
this figure show That The alignment plot scores higher diagonal intensity than MCB-MS2 plot which indicates higher protein stability in the presence of NB1 .
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4
Illegal EcoRI site found at 282 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4
Illegal EcoRI site found at 282 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4
Illegal EcoRI site found at 282 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4
Illegal EcoRI site found at 282 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4
Illegal EcoRI site found at 282 - 1000COMPATIBLE WITH RFC[1000]
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