He M, Jin L, Austen B
Department of Surgery, St. George's Hospital Medical School, Tooting, London, United Kingdom.
J Protein Chem. 1993 Feb;12(1):1-5. doi: 10.1007/BF01024906.
The precursor protein honey bee prepromelittin has been expressed as a fusion protein in Escherichia 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.
蜜蜂前催乳素原蛋白已在大肠杆菌中作为融合蛋白表达，该融合蛋白通过针对凝血因子Xa的工程识别序列与噬菌体基因10蛋白截短形式的C末端相连。发现凝血因子Xa在工程位点的切割效果不佳，所需前催乳素原蛋白的产量较低。相比之下，在基因10序列中第67位残基处VLGR序列的C末端一侧进行切割时，产量较高。凝血因子Xa可能会受到潜在切割位点C末端一侧相邻疏水序列的抑制。
