Lebherz H G, Doyle D
J Biol Chem. 1976 Jul 25;251(14):4355-8.
The present report describes the complete synthesis of a functional oligomeric enzyme in a heterologous cell-free system. Polysomal RNA from chicken skeletal muscle was used to direct the production of functional aldolase tetramers in wheat germ extracts. The aldolase product was (a) specifically precipitated with monospecific antibodies raised against pure muscle aldolase, (b) had the same subunit molecular weight (40,000) as that of native aldolase (as determined by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate), (c) presumably contained a functional active site since it co-purified with authentic muscle aldolase upon substrate elution from phosphocellulose, and (d) had associated into tetrameric units (Mr=160,000) as shown by centrifugation in sucrose gradients. The present work suggests that, within the cell, post-translational processing of aldolase polypeptide chains is not involved in the formation of functional aldolase tetramers.
本报告描述了在异源无细胞系统中功能性寡聚酶的完全合成。来自鸡骨骼肌的多聚核糖体RNA用于指导在小麦胚芽提取物中产生功能性醛缩酶四聚体。醛缩酶产物：(a) 被针对纯肌肉醛缩酶产生的单特异性抗体特异性沉淀；(b) 具有与天然醛缩酶相同的亚基分子量（40,000）（通过在十二烷基硫酸钠存在下的丙烯酰胺凝胶电泳测定）；(c) 可能含有功能性活性位点，因为在从磷酸纤维素上用底物洗脱时，它与真实的肌肉醛缩酶共纯化；(d) 如在蔗糖梯度中离心所示，已缔合成四聚体单元（Mr = 160,000）。目前的工作表明，在细胞内，醛缩酶多肽链的翻译后加工不参与功能性醛缩酶四聚体的形成。
