Lee B, Tabita F R
Department of Microbiology, Ohio State University, Columbus 43210.
Biochemistry. 1990 Oct 9;29(40):9352-7. doi: 10.1021/bi00492a007.
Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) from Anacystis nidulans was reconstituted in vitro from extracts of Escherichia coli strains that separately express large and small subunits. This reconstitution system was shown to be useful for monitoring the appearance of dissociated or fractionated subunit preparations. Recombinant large subunits were purified to a state of homogeneity and retained reconstitution capacity in the presence of added small subunits. The purified large subunits appeared to be in the form of an octamer, probably an L8 structure, and showed 0.15% of the carboxylase activity of the purified L8S8 enzyme. Purified large subunit octamers are disrupted by nondenaturing PAGE; however, the octamer is stable to electrophoresis in the presence of exogenous protein.
来自集胞藻6803的1,5-二磷酸核酮糖羧化酶/加氧酶（RubisCO）在体外由分别表达大亚基和小亚基的大肠杆菌菌株提取物重构而成。该重构系统被证明可用于监测解离或分级分离的亚基制剂的出现情况。重组大亚基被纯化至同质状态，并在添加小亚基的情况下保留重构能力。纯化的大亚基似乎以八聚体形式存在，可能是L8结构，并且显示出纯化的L8S8酶羧化酶活性的0.15%。纯化的大亚基八聚体可通过非变性聚丙烯酰胺凝胶电泳（PAGE）破坏；然而，在存在外源蛋白的情况下，八聚体对电泳是稳定的。
