Taubman M A, Smith D J, King W F, Eastcott J W, Bergey E J, Levine M J
Department of Immunology, Forsyth Dental Center, Boston, MA 02115.
J Oral Pathol. 1988 Nov;17(9-10):466-70. doi: 10.1111/j.1600-0714.1988.tb01317.x.
GTF activity was separated into water-insoluble (GTF-I) and water-soluble (GTF-S) polyglucan-synthesizing enzymes. Each preparation demonstrated a single band on 6% SDS PAGE. Only water-insoluble or water-soluble polyglucan was synthesized by the respective enzyme preparation. Rats were immunized, on Days 1 and 14, with either GTF-I or GTF-S in adjuvant. Animals were bled 13, 35 and 54 days after the initial immunization. Individual antisera were tested against either the GTF-I or the GTF-S for inhibition of radioactive glucose incorporation into glucan, and in gel diffusion, and by Western transfer analyses. The respective antisera reacted with the homologous, but not the heterologous enzyme in gel diffusion and Western transfer. GTF-I activity was not inhibited by antibody to GTF-S, but antibody to GTF-I inhibited GTF-I by 68%. GTF-S was inhibited by more than 60% by each of 3 anti-GTF-S sera. Only one anti-GTF-I serum inhibited GTF-S at as much as a modest 30% level. These data support the antigenic and functional distinctiveness of the GTF enzymes of S. sobrinus 6715.
葡糖基转移酶（GTF）活性被分离为水不溶性（GTF-I）和水溶性（GTF-S）的多聚糖合成酶。每种制剂在6%十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS PAGE）上显示出一条带。相应的酶制剂仅合成水不溶性或水溶性多聚糖。在第1天和第14天，用佐剂中的GTF-I或GTF-S对大鼠进行免疫。在初次免疫后13、35和54天采集动物血液。针对GTF-I或GTF-S检测个体抗血清对放射性葡萄糖掺入葡聚糖的抑制作用、凝胶扩散以及蛋白质免疫印迹分析。在凝胶扩散和蛋白质免疫印迹中，相应的抗血清与同源酶反应，但不与异源酶反应。GTF-S抗体不抑制GTF-I活性，但GTF-I抗体抑制GTF-I活性达68%。3种抗GTF-S血清中的每一种都使GTF-S受到超过60%的抑制。只有一种抗GTF-I血清对GTF-S的抑制作用可达适度的30%水平。这些数据支持了远缘链球菌6715的GTF酶在抗原性和功能上的独特性。
