Ganesa C, Flurkey W H, Randhawa Z I, Bozarth R F
Department of Life Sciences, Indiana State University, Terre Haute 47809.
Arch Biochem Biophys. 1991 Apr;286(1):195-200. doi: 10.1016/0003-9861(91)90027-g.
The toxin from Ustilago maydis virus P4 was purified to homogeneity and characterized. The native molecular mass, using size-exclusion HPLC was estimated to be 7.2 kDa. The purified toxin was composed of a single subunit. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis under reduced and nonreduced conditions resulted in estimated molecular masses of 8.4 and 7.4 kDa, respectively. The purified toxin was found to be glycosylated when tested for carbohydrates using the phenol-sulfuric acid method, Schiff's base reagent, and a Glycan detection kit and when probed against different biotinylated lectins. Partial amino acid sequence analysis of the purified toxin indicated a free N-terminus, 16% glycine, and 23% basic amino acid residues. No homology was found to either the alpha or the beta subunit of the toxin encoded by U. maydis infected with the P6 virus.
对玉米黑粉菌病毒P4的毒素进行了纯化，直至达到同质，并对其进行了特性鉴定。使用尺寸排阻高效液相色谱法估计其天然分子量为7.2 kDa。纯化后的毒素由单个亚基组成。在还原和非还原条件下进行的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析得出的估计分子量分别为8.4 kDa和7.4 kDa。当使用苯酚-硫酸法、席夫碱试剂和聚糖检测试剂盒检测碳水化合物，并与不同的生物素化凝集素进行检测时，发现纯化后的毒素是糖基化的。对纯化毒素的部分氨基酸序列分析表明其N端游离，含有16%的甘氨酸和23%的碱性氨基酸残基。未发现与感染P6病毒的玉米黑粉菌编码的毒素的α或β亚基有同源性。
