Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271-8587, Japan.
Microb Pathog. 2010 Mar-Apr;48(3-4):116-23. doi: 10.1016/j.micpath.2010.01.001. Epub 2010 Jan 11.
Hemin-binding protein 35 (HBP35) may be an essential protein for bacterial survival in evasion from environmental stress in Porphyromonas gingivalis. The anti-recombinant HBP35 antibody inhibits P. gingivalis hemagglutination. This study considered the role of this protein for hemagglutination and adherence to host cells using the HBP35-deficient mutant (MD774) derived from P. gingivalis FDC381. FDC381 had strong hemagglutination activity, whereas MD774 had no activity. Anti-130-kDa hemagglutinin antibody, mAb-Pg-vc, reacted mainly with the 43- and 49-kDa molecules in the membrane fraction. However, no proteins reacted in the MD774. The hemolytic activity in MD774 was much lower than that in FDC381. Anti-recombinant HBP35 antibody strongly inhibited the P. gingivalis FDC381 adherence to epithelial cells. In addition, MD774 exhibited a significant decrease in the adherence. The hydrophobicity of MD774 was equal to 19.4% of that of FDC381. SDS-PAGE profiling of the membrane fractions of both strains showed very different profiles. Taken together, these findings suggest that HBP35 plays a role, not only in hemin-binding, but also in multiple P. gingivalis binding to erythrocytes, and host epithelial gingival cells. In addition, this protein may directly and/or indirectly affect the virulence of this organism.
血红素结合蛋白 35（HBP35）可能是牙龈卟啉单胞菌逃避环境压力时生存的必需蛋白。抗重组 HBP35 抗体抑制牙龈卟啉单胞菌的红细胞凝集作用。本研究利用源自牙龈卟啉单胞菌 FDC381 的 HBP35 缺陷突变株（MD774），研究了该蛋白在红细胞凝集和黏附宿主细胞中的作用。FDC381 具有很强的红细胞凝集活性，而 MD774 则没有活性。抗 130-kDa 血凝素抗体 mAb-Pg-vc 主要与膜部分的 43-和 49-kDa 分子反应。然而，MD774 则没有蛋白质反应。MD774 的溶血活性明显低于 FDC381。抗重组 HBP35 抗体强烈抑制牙龈卟啉单胞菌 FDC381 对上皮细胞的黏附。此外，MD774 的黏附作用显著下降。MD774 的疏水性为 FDC381 的 19.4%。两种菌株的膜部分 SDS-PAGE 分析显示出非常不同的图谱。总之，这些发现表明 HBP35 不仅在血红素结合中发挥作用，而且在牙龈卟啉单胞菌与红细胞和宿主上皮牙龈细胞的多种结合中也发挥作用。此外，该蛋白可能直接和/或间接影响该生物体的毒力。
