pMCP-tag(m) Vector is a mammalian expression plasmid encoding MCP, an MCP-tag protein, which is expressed under control of the CMV promoter. This plasmid is intended for the cloning and transient expression of MCP-tag protein fusions in mammalian cells. pMCP-tag(m) Vector contains two multiple cloning sites to allow cloning of the fusion partner as a fusion to the C-terminus of the MCP-tag and an appropriate signal peptide to the N-terminus of the MCP-tag.
The MCP-tag is a small protein tag (8 kDa) based on the acyl carrier protein (ACP) containing two mutations (D36-T36 and D39-G39), for labeling cell membrane proteins. It allows the specific, covalent attachment of virtually any molecule to a protein of interest. MCP-tag substrates are derivatives of coenzyme A (CoA). In the labeling reaction, the substituted phosphopantetheine group of CoA is covalently attached to a conserved serine residue of the MCP-tag by SFP Synthase. Having no cysteines, the MCP-tag is particularly suited for specifically labeling cell-surface proteins, and should be useful for labeling secreted proteins with disulfide bridges such as antibodies.
There are two steps to using this system: sub cloning and expression of the protein of interest as a MCP-tag fusion, and labeling of the fusion protein using SFP Synthase with the substrate of choice. The cloning and expression of MCP-tag protein fusions is described in this document. The labeling of fusion proteins with CoA substrates is described in the documentation supplied with CoA substrates and SFP Synthase.
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