Brunngraber E F, Chargaff E
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3226-9. doi: 10.1073/pnas.74.8.3226.
Improved extraction and purification procedures permit the isolation from Escherichia coli W cells of much larger quantities and of more highly purified preparations of nucleotide phosphotransferase. Of various affinity resins tested for efficiency of purification, columns of agarose/5'-AMP (AGAMP), type 3, proved the best. In this way a 300- to 450-fold purification of the enzyme was achieved in a few steps. The enzyme, which, as reported before, transfers organically bound phosphate to the 2' or 3' hydroxyls of nucleosides and nucleotides, was tested in its behavior toward a series of ribonucleosidonucleotides, namely, CpC, ApA, CpA, and ApC. All were phosphate acceptors, but a detailed comparative study of adenosine and cytidine, 5'-AMP and 5'-CMP, and ApA and ApC revealed peculiar specificities in the relative distribution of the phosphorylated products.
改进的提取和纯化程序能够从大肠杆菌W细胞中分离出数量更多、纯度更高的核苷酸磷酸转移酶制剂。在测试的各种用于纯化效率的亲和树脂中，3型琼脂糖/5'-AMP（AGAMP）柱被证明是最好的。通过这种方法，在几步之内就实现了该酶300至450倍的纯化。如之前所报道的，该酶将有机结合的磷酸转移到核苷和核苷酸的2'或3'羟基上，对一系列核糖核苷核苷酸，即CpC、ApA、CpA和ApC的行为进行了测试。它们都是磷酸受体，但对腺苷和胞苷、5'-AMP和5'-CMP以及ApA和ApC的详细比较研究揭示了磷酸化产物相对分布中的特殊特异性。
