PURIFICATION AND CHARACTERIZATION OF 5'-DEOXY-5'-METHYLTHIOADENOSINE (MTA) PHOSPHORYLASE FROM HUMAN LIVER
PURIFICATION AND CHARACTERIZATION OF 5'-DEOXY-5'-METHYLTHIOADENOSINE (MTA) PHOSPHORYLASE FROM HUMAN LIVER
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DOI：
10.1016/0006-2952(91)90145-u
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发表时间：
1991-06-15
影响因子：
5.8
通讯作者：
MILLER, RL
中科院分区：
文献类型：
--
作者：
TOORCHEN, D;MILLER, RL
使用亲和层析、层析聚焦和凝胶过滤相结合，从人肝脏中纯化 5'-甲硫腺苷磷酸化酶 8000 倍。获得25%的总产率。最终制剂的比活性为每小时每毫克蛋白质裂解 40-μmol 5'-甲硫腺苷。通过Superose 12和Sephadex G-150凝胶过滤测定，该酶的表观分子量为55,000道尔顿，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定，亚基分子量为30,000道尔顿。在核苷裂解和核苷合成两个方向上研究了纯化酶的底物特异性。
5'-Methylthioadenosine phosphorylase was purified 8000-fold from human liver using a combination of affinity chromatography, chromatofocusing and gel filtration. A 25% overall yield was obtained. The specific activity of the final preparation was 40-mu-mol of 5'-methylthioadenosine cleaved per hr per mg of protein. The enzyme had an apparent molecular weight of 55,000 daltons, as determined by gel filtration on Superose 12 and Sephadex G-150, with a subunit molecular weight of 30,000 daltons, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The substrate specificity of the purified enzyme was studied in both the direction of nucleoside cleavage and nucleoside synthesis.