Takai I, Oda O, Shinzato T, Ohbayashi K, Yamanaka N, Maeda K
Department of Internal Medicine, Branch Hospital, Nagoya University School of Medicine, Japan.
J Chromatogr B Biomed Appl. 1996 Oct 11;685(1):21-5. doi: 10.1016/0378-4347(96)00161-2.
Lysozyme in the urine of a hemodialysis patient was purified in two steps: DEAE Sephadex chromatography followed by Sephacryl chromatography. The Sephacryl S-100 column chromatographed fraction showing lytic activity was proven to give one band on SDS-PAGE and to have a molecular mass of 14500, in agreement with that of lysozyme. The N-terminal amino acid sequence of this purified protein was identical to that of lysozyme. These results indicate that the protein purified was indeed lysozyme. The specific affinity of lysozyme for Sephacryl S-100 may explain the greater purity of the same protein isolated by this method.
首先进行DEAE Sephadex柱色谱，然后进行Sephacryl柱色谱。经Sephacryl S - 100柱色谱分离得到的具有溶菌活性的组分，在SDS - PAGE上显示为一条带，其分子量为14500，与溶菌酶一致。该纯化蛋白的N端氨基酸序列与溶菌酶相同。这些结果表明纯化得到的蛋白确实是溶菌酶。溶菌酶对Sephacryl S - 100的特异性亲和力可能解释了用该方法分离得到的同一蛋白具有更高纯度的原因。
