Ito K
J Biol Chem. 1982 Sep 10;257(17):9895-7.
Cells of Escherichia coli K12 strain MM18 accumulate precursors of various exported proteins when the synthesis of the malE-lacZ hybrid protein is induced by maltose. Starting from a soluble cell extract of this strain, the precursor form of the maltose-binding protein, a periplasmic protein, was purified to homogeneity. The procedure includes affinity chromatography on cross-linked amylose and chromatofocusing. The purified precursor has an intact signal sequence at the NH2 terminus. It is soluble in aqueous buffer probably in monomeric state and is proteolytically processed by the leader peptidase, which was previously purified by Wickner and co-workers as an enzyme that processes M13 phage procoat protein.
当麦芽糖诱导麦芽糖结合蛋白(MalE)-乳糖酶(LacZ)杂合蛋白合成时,大肠杆菌K12菌株MM18的细胞会积累各种输出蛋白的前体。从该菌株的可溶性细胞提取物开始,将周质蛋白麦芽糖结合蛋白的前体形式纯化至同质。该方法包括在交联直链淀粉上进行亲和色谱和色谱聚焦。纯化的前体在NH2末端具有完整的信号序列。它可能以单体状态溶于水性缓冲液中,并被前导肽酶进行蛋白水解加工,前导肽酶先前由维克纳及其同事纯化,作为一种加工M13噬菌体原衣壳蛋白的酶。