Specificity of Factor Xa in the cleavage of fusion proteins |
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Authors: | Mingyue He Li Jin and Brian Austen |
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Institution: | (1) Peptide Unit, Department of Surgery, St. George's Hospital Medical School, Tooting, SW17 0RE London, United Kingdom |
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Abstract: | The precursor protein honey bee prepromelittin has been expressed as a fusion protein inEscherichia coli joined to the C-terminus of a truncated form of the bacteriophage gene 10 protein via an engineered recognition sequence for Factor Xa. Factor Xa was found to cleave poorly at the engineered site, giving a low yield of the required prepromelittin. In contrast, cleavage on the C-terminal side of the sequence VLGR at residue 67 in the gene 10 sequence proceeded in high yield. Factor Xa may be inhibited by adjacent hydrophobic sequences on the C-terminal side of a potential cleavage site. |
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Keywords: | Factor Xa fusion proteins bacterial expression proteolytic specificity |
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