Gauthier L, Mayrand D, Vadeboncoeur C
J Bacteriol. 1984 Nov;160(2):755-63. doi: 10.1128/jb.160.2.755-763.1984.
Fructose transport in Streptococcus mutans LG-1 is mediated by at least two distinct phosphoenolpyruvate fructose phosphotransferase systems. One system is constitutive and consists of membrane components enzyme II as well as enzyme I and heat-stable protein. The other system is inducible and requires, in addition to enzyme I and heat-stable protein, a soluble substrate-specific protein for catalytic activity. This protein factor, designated IIIfru, was purified by DEAE-cellulose chromatography, hydroxylapatite chromatography, molecular sieving on Sephadex G-75, and preparative electrophoresis. The purified preparation showed only one protein band, with a molecular weight of 12,600, on sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis, on gel electrophoresis with the discontinuous buffer Tris-glycine, and after electrofocusing in gel (pI congruent to 3.7). The molecular weight of the native protein determined by gel filtration at 4 degrees C was 51,000. Immunodiffusion experiments performed with immunoglobulins prepared against the purified IIIfru from S. mutans LG-1 suggested that other S. mutans strains possessed a IIIfru. No precipitin bands, however, were detected with extracts from S. salivarius, S. sanguis, S. lactis, S. faecalis, Staphylococcus aureus, Bacillus subtilis, Lactobacillus casei, and Escherichia coli.
变形链球菌LG-1中的果糖转运至少由两种不同的磷酸烯醇丙酮酸-果糖磷酸转移酶系统介导。一种系统是组成型的,由膜成分酶II以及酶I和热稳定蛋白组成。另一种系统是诱导型的,除酶I和热稳定蛋白外,还需要一种可溶性底物特异性蛋白来发挥催化活性。这种蛋白因子,命名为IIIfru,通过DEAE-纤维素色谱法、羟基磷灰石色谱法、Sephadex G-75分子筛和制备电泳进行纯化。在十二烷基硫酸钠-尿素-聚丙烯酰胺凝胶电泳、使用不连续缓冲液Tris-甘氨酸的凝胶电泳以及在凝胶中进行聚焦电泳后(pI等于3.7),纯化制剂在凝胶上仅显示一条分子量为12,600的蛋白带。在4℃下通过凝胶过滤测定的天然蛋白分子量为51,000。用针对变形链球菌LG-1纯化的IIIfru制备的免疫球蛋白进行的免疫扩散实验表明,其他变形链球菌菌株也拥有IIIfru。然而,用唾液链球菌、血链球菌、乳酸链球菌、粪肠球菌、金黄色葡萄球菌、枯草芽孢杆菌、干酪乳杆菌和大肠杆菌的提取物未检测到沉淀带。