Weiss E I, Kolenbrander P E, London J, Hand A R, Andersen R N
J Bacteriol. 1987 Sep;169(9):4215-22. doi: 10.1128/jb.169.9.4215-4222.1987.
Bacteroides loescheii PK1295 serves as a coaggregation bridge between Streptococcus sanguis 34 and Actinomyces israelii PK14, two gram-positive oral bacteria that are otherwise unable to coaggregate. Whereas coaggregation with S. sanguis 34 is inhibited by lactose, no simple sugar was found that inhibited coaggregation with A. israelii PK14. Coaggregation-defective (Cog-) mutants of B. loescheii PK1295 were isolated for the purpose of identifying the surface components responsible for the interaction with each coaggregation partner. Selection for spontaneously occurring Cog- mutants gave rise to two phenotypic classes of mutants. Type I lost the ability to coaggregate with S. sanguis 34, whereas type II failed to coaggregate with either S. sanguis 34 or A. israelii PK14. Purified fimbriae from the parent agglutinated cells of both partners, and agglutination with S. sanguis 34 was inhibited by lactose. Denaturing polyacrylamide gel electrophoresis and immunoblot analysis demonstrated the presence of both a 75- and a 43-kilodalton (kDa) protein associated with parental fimbriae, but only a 43-kDa protein was seen with fimbriae prepared from the type I mutant. Neither polypeptide was found in similar preparations from the type II mutants. Our data suggest that coaggregation of B. loescheii PK1295 with both gram-positive partners is mediated by fimbria-associated proteins present on the surface of the gram-negative organism and that the 75- and 43-kDa polypeptides are responsible for the recognition of S. sanguis 34 and A. israelii PK14 cells, respectively.
洛氏拟杆菌PK1295在血链球菌34和以色列放线菌PK14之间起到共聚集桥梁的作用,这两种革兰氏阳性口腔细菌在其他情况下无法发生共聚集。与血链球菌34的共聚集受到乳糖的抑制,但未发现能抑制与以色列放线菌PK14共聚集的单糖。为了确定与每个共聚集伙伴相互作用的表面成分,分离出了洛氏拟杆菌PK1295的共聚集缺陷(Cog-)突变体。对自发产生的Cog-突变体进行筛选,得到了两类表型突变体。I型失去了与血链球菌34共聚集的能力,而II型则无法与血链球菌34或以色列放线菌PK14共聚集。来自亲本的纯化菌毛能凝集两种伙伴的细胞,与血链球菌34的凝集受到乳糖的抑制。变性聚丙烯酰胺凝胶电泳和免疫印迹分析表明,与亲本菌毛相关的有一个75千道尔顿(kDa)和一个43 kDa的蛋白质,但从I型突变体制备的菌毛中只看到了一个43 kDa的蛋白质。在II型突变体的类似制剂中未发现这两种多肽。我们的数据表明,洛氏拟杆菌PK1295与这两种革兰氏阳性伙伴的共聚集是由革兰氏阴性生物体表面存在的菌毛相关蛋白介导的,并且75 kDa和43 kDa的多肽分别负责识别血链球菌34和以色列放线菌PK14细胞。