Layh-Schmitt G, Schmitt S, Buchanan T M
Department of Pathobiology, University of Washington, Seattle 98195.
Zentralbl Bakteriol. 1989 Jul;271(2):158-70. doi: 10.1016/s0934-8840(89)80069-6.
Studies with [14C] uracil-labelled bacteria revealed that the interaction of Neisseria gonorrhoeae with epithelial cells occurred in a time-dependent reaction which is slightly pH-dependent and optimal at pH 6.5. Immunofluorescence tests and immunoelectron microscopy of ultrathin sections confirmed the attachment of these bacteria to the epithelial cell membrane. The interaction of purified protein I with epithelial cells was time-dependent and reached equilibrium after four hours as shown by tracer experiments with 125I-labeled protein I. Cleavage experiments with trypsin followed by SDS-PAGE and autoradiography indicated that protein I (labeled with 125I) was associated with the membrane of the epithelial cells and only partly accessible by trypsin after its interaction with these mammalian cells. Immunofluorescence tests as well as immunoelectron microscopy with the monoclonal antibody G7A2C and gold-labeled protein A confirmed a dense association pattern of protein I with the cell monolayer.
用[14C]尿嘧啶标记细菌的研究表明,淋病奈瑟菌与上皮细胞的相互作用发生在一个时间依赖性反应中,该反应略受pH值影响,在pH 6.5时最为适宜。超薄切片的免疫荧光试验和免疫电子显微镜检查证实了这些细菌附着在上皮细胞膜上。如用125I标记蛋白I的示踪实验所示,纯化蛋白I与上皮细胞的相互作用是时间依赖性的,4小时后达到平衡。用胰蛋白酶进行切割实验,随后进行SDS-PAGE和放射自显影,结果表明蛋白I(用125I标记)与上皮细胞膜相关,在与这些哺乳动物细胞相互作用后,胰蛋白酶只能部分作用于它。免疫荧光试验以及用单克隆抗体G7A2C和金标蛋白A进行的免疫电子显微镜检查证实了蛋白I与细胞单层的密集结合模式。