MECHANISMS OF PORPHYROMONAS/STREPTOCOCCUS COHESION

卟啉单胞菌/链球菌凝聚机制

• 批准号: 6296259
• 负责人: MURRAY W STINSON
• 金额: $ 14.59万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6296259
• 关键词: Bacteroides gingivalis; SDS polyacrylamide gel electrophoresis; Streptococcus sanguis; adhesions; binding proteins; cell adhesion; cell membrane; dental plaque; disease /disorder model; high performance liquid chromatography; human subject; ion exchange chromatography; laboratory rabbit; laboratory rat; membrane proteins; microorganism culture; microorganism interaction; molecular cloning; nucleic acid sequence; pathologic bone resorption; pellicle; periodontium disorder; protein purification; proteolysis; receptor binding; saliva; serum; site directed mutagenesis; structural genes; surface antigens; virulence; western blottings

The objective of this subproject is to further characterize the mechanism(s) involved in cohesion between Porphyromonas gingivalis and Streptococcus gordonii or Streptococcus sanguis. In particular, the mechanisms whereby selected saliva and serum components inhibit this cohesion will be explored. Our previous studies have demonstrated that outer membrane proteins (OMP) of P. gingivalis may be involved in these interactions. Consequently, outer membrane adhesins of P. gingivalis will be identified in binding assays using radiolabeled ligand molecules from saliva or streptococci. The adhesins will be purified using a combination of gel filtration, ion exchange, and affinity chromatography. The N- terminal sequence of the purified protein will be determined and used to deduce the nucleotide sequence of this region of the gene. Synthetic oligonucleotide probes will be used to identify the structural genes, which will be isolated, cloned, and the complete nucleotide sequences determined. Adhesin-deficient strains of P. gingivalis will be generated by transposon insertion mutagenesis and tested for their abilities to adhere to experimental streptococcal plaque and experimental salivary pellicles. The surface composition of adhesin-deficient mutants will be compared with those of isogeneic parental strains to identify other possible adhesins. We anticipate that the information derived from these studies will provide a rationale for enhancing natural host defenses to minimize colonization of dental plaque and the oral cavity by P. gingivalis and related oral pathogens.

该子项目的目标是进一步表征 牙龈卟啉单胞菌之间的凝聚机制 戈登链球菌或血链球菌。 特别是， 选定的唾液和血清成分抑制这种作用的机制 将探讨凝聚力。 我们之前的研究表明 牙龈卟啉单胞菌的外膜蛋白（OMP）可能参与这些过程 互动。 因此，牙龈卟啉单胞菌的外膜粘附素将 使用放射性标记的配体分子在结合测定中进行鉴定 唾液或链球菌。 粘附素将使用组合来纯化 凝胶过滤、离子交换和亲和层析。 然后- 纯化蛋白的末端序列将被确定并用于 推导出该基因区域的核苷酸序列。 合成的 寡核苷酸探针将用于鉴定结构基因， 将被分离、克隆并确定完整的核苷酸序列。 转座子将产生粘附素缺陷型牙龈卟啉单胞菌菌株 插入诱变并测试其粘附能力 实验性链球菌菌斑和实验性唾液膜。 这 粘附素缺陷突变体的表面组成将与 那些同基因亲本菌株，以鉴定其他可能的粘附素。 我们预计从这些研究中获得的信息将提供 增强自然宿主防御以尽量减少定植的理由 牙龈卟啉单胞菌和相关口腔细菌对牙菌斑和口腔的影响 病原体。