Hackett M, Walker C B, Guo L, Gray M C, Van Cuyk S, Ullmann A, Shabanowitz J, Hunt D F, Hewlett E L, Sebo P
Department of Chemistry, University of Virginia, Charlottesville 22901, USA.
J Biol Chem. 1995 Sep 1;270(35):20250-3. doi: 10.1074/jbc.270.35.20250.
Adenylate cyclase toxin from Bordetella pertussis requires posttranslational acylation of lysine 983 for the ability to deliver its catalytic domain to the target cell interior and produce cyclic adenosine monophosphate (cell-invasive activity) and to form transmembrane channels (hemolytic activity). When the toxin is expressed in Escherichia coli, it has reduced hemolytic activity, but comparable cell-invasive activity to that of adenylate cyclase toxin from B. pertussis. In contrast to the native protein from B. pertussis, which is exclusively palmitoylated, recombinant toxin from E. coli is acylated at lysine 983 with about 87% palmitoylated and the remainder myristoylated. Furthermore, the recombinant toxin contains an additional palmitoylation on approximately two-thirds of the lysines at position 860. These observations suggest that the site and nature of posttranslational fatty-acylation can be dictated by the bacterial host used for expression and can have a significant, but selective, effect on protein function.
百日咳博德特氏菌的腺苷酸环化酶毒素需要赖氨酸983进行翻译后酰化，才能将其催化结构域递送至靶细胞内部并产生环磷酸腺苷（细胞侵袭活性），以及形成跨膜通道（溶血活性）。当该毒素在大肠杆菌中表达时，其溶血活性降低，但细胞侵袭活性与百日咳博德特氏菌的腺苷酸环化酶毒素相当。与仅被棕榈酰化的百日咳博德特氏菌天然蛋白不同，来自大肠杆菌的重组毒素在赖氨酸983处被酰化，约87%为棕榈酰化，其余为肉豆蔻酰化。此外，重组毒素在约三分之二位于860位的赖氨酸上还存在额外的棕榈酰化。这些观察结果表明，翻译后脂肪酰化的位点和性质可由用于表达的细菌宿主决定，并且对蛋白质功能可产生显著但具有选择性的影响。
