Part:BBa_K1766012:Design
Z:HER2:342 for GPI anchoring
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 397
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 24
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 299
Design Notes
This is a design for a membrane-bound Affibody molecule with affinity to the cancer protein HER-2.
Our design used a system of GPI-anchoring common in membrane proteins of mammalian cells. We based our design on Human protein DAF, encoded by the CD55 gene. A 34 amino acid long signal peptide at the N terminal precedes the actual protein, which is then followed by a second 28 amino acid long signal peptide at the C terminal. The N-terminal signal directs the protein to the ER, and the C-terminal signal is removed in the ER and is replaced with a GPI anchor. Addition of a DAF-derived signal has been shown to lead to GPI anchoring of fusion proteins.
In addition to the signal peptide, we also added a short linker sequence and a FLAG-tag on the N-terminal of the Affibody. On the C-terminal, we also added a linker that ended with Serine, which is the site of cleavage for the signal peptide.
Source
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