Russell G C, Williamson R A, Guest J R
Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, U.K.
FEMS Microbiol Lett. 1989 Aug;51(3):267-71. doi: 10.1016/0378-1097(89)90408-4.
Two compatible plasmids encoding a hybrid lipoyl domain and a defective pyruvate dehydrogenase (PDH) complex which lacks lipoyl domains, were co-expressed in a strain of Escherichia coli deleted for the PDH complex genes. In vivo complementation between the mutant complexes and the independent lipoyl domains was observed using growth tests in liquid and solid media. However, no PDH complex activity could be detected in the corresponding cell-free extracts. This suggests that untethered lipoyl domains can interact productively with the three types of active site in the multienzyme complex, but this association is disrupted in cell-free extracts.
两个相容的质粒，一个编码杂合硫辛酰结构域，另一个编码缺乏硫辛酰结构域的缺陷型丙酮酸脱氢酶（PDH）复合物，在缺失PDH复合物基因的大肠杆菌菌株中共同表达。通过在液体和固体培养基中进行生长测试，观察到突变复合物与独立硫辛酰结构域之间的体内互补作用。然而，在相应的无细胞提取物中未检测到PDH复合物活性。这表明未连接的硫辛酰结构域可以与多酶复合物中的三种活性位点有效相互作用，但这种结合在无细胞提取物中被破坏。
