In the case of the C-terminal fusion vectors (pTYB1,2,3 and 4), which residues at the C-terminal of the target protein may inhibit cleavage or cause in vivo cleavage?
The data in the table below is based on analysis of cleavage reactions using the E. coli maltose-binding protein (MBP) as the target protein (MYB) with amino acid substitutions at the position (-1) immediately upstream of the cleavage site in the sequence (L-3E-2 X-1/C1). In summary, the data indicate that Pro, Cys and Asn inhibit thiol-induced cleavage, while Asp, Arg, His, Glu and Thr cause substantial in vivo cleavage of the fusion proteins. Therefore if the target protein contains one of these “unfavorable residues” at its C-terminus, the inclusion of a favorable residue or sequence immediately adjacent to the cleavage site may be necessary for controllable cleavage reaction. In the case of the MBP fusion, when a glycine is added between the proline and the intein N-terminal cysteine, inducible cleavage is greatly improved. However, this is not to say that all proteins will behave in the same manner when fused to the intein-chitin binding domain. In each case, the effect of differe
Related Questions
- In the case of the C-terminal fusion vectors (pTYB1,2,3 and 4), which residues at the C-terminal of the target protein may inhibit cleavage or cause in vivo cleavage?
- In the case of the N-terminal fusion vectors (pTYB11 and 12), which residues at the N-terminal of the target protein may inhibit cleavage or cause in vivo cleavage?
- Which residues at the C-terminus of the target protein may inhibit cleavage or cause in vivo cleavage when the pTXB vectors are used?
