Gibbons R J, Hay D I
Department of Microbiology, Forsyth Dental Center, Boston, Massachusetts 02115.
Infect Immun. 1988 Feb;56(2):439-45. doi: 10.1128/iai.56.2.439-445.1988.
Actinomyces viscosus LY7 cells adsorbed in high numbers to experimental pellicles formed on hydroxyapatite (HA) from human parotid or submandibular saliva but not to pellicles prepared from human plasma or serum. To determine the nature of the salivary components responsible for promoting adhesion, pellicles were prepared from fractions of submandibular and parotid saliva obtained by chromatography on Trisacryl GF 2000 columns. Adsorption of LY7 cells was promoted by two groups of fractions. Each group was rechromatographed on DEAE-agarose. Fractions which promoted adsorption of LY7 cells were found by polyacrylamide gel electrophoresis to contain the acidic proline-rich proteins (PRPs) and statherin. Pellicles prepared from 12-micrograms/ml solutions of pure PRP-1, PRP-2, or parotid isoelectric focusing (PIF-slow) variant promoted maximal adsorption of A. viscosus LY7 cells. Somewhat higher concentrations of PRP-3 and PRP-4 were required for maximal adsorption, indicating that the 44-residue carboxy-terminal segment of PRP-1, PRP-2, and PIF-slow enhances LY7 binding but is not essential. Much higher concentrations of statherin were required to promote LY7 adsorption. Adsorption of LY7 cells to pellicles prepared from PRP-1 was not affected over the range of pH 5 to 8. Adsorption was also not inhibited by 50 mM lactose, which is consistent with the notion that type 1 fimbriae, rather than type 2 fimbriae, were responsible. A. viscosus T14, Actinomyces odontolyticus ATCC 17982, and Actinomyces israelii 12597 also adsorbed to PRP-1 pellicles, whereas Actinomyces naeslundii ATCC 12104 did not. Although A. viscosus cells bind strongly to adsorbed PRP-1, the presence of PRP-1 or PRP-3 in solution did not inhibit adhesion. Similarly, [3H]PRP-1 did not bind to LY7 cells, nor was it degraded when incubated with the organism. However, LY7 cells adsorbed to [3H]PRP-1 pellicles. These data suggest that hidden molecular segments of PRP become exposed when the protein adsorbs to HA; these segments then react with adhesins of LY7 cells. The apparent ability of A. viscosus cells to recognize segments of PRPs which are exposed only in surface-adsorbed molecules provides a novel mechanism which enables the organism to attach to teeth when suspended in salivary secretions.
黏性放线菌LY7细胞大量吸附于由人腮腺或下颌下腺唾液在羟基磷灰石(HA)上形成的实验性薄膜,但不吸附于由人血浆或血清制备的薄膜。为确定促进黏附的唾液成分的性质,用在Trisacryl GF 2000柱上进行色谱分离得到的下颌下腺和腮腺唾液组分制备薄膜。两组组分促进了LY7细胞的吸附。每组在DEAE -琼脂糖上再次进行色谱分离。通过聚丙烯酰胺凝胶电泳发现,促进LY7细胞吸附的组分含有酸性富含脯氨酸蛋白(PRPs)和磷蛋白。由12微克/毫升的纯PRP - 1、PRP - 2或腮腺等电聚焦(PIF -慢)变体溶液制备的薄膜促进了黏性放线菌LY7细胞的最大吸附。PRP - 3和PRP - 4需要稍高的浓度才能实现最大吸附,这表明PRP - 1、PRP - 2和PIF -慢的44个残基的羧基末端片段增强了LY7的结合,但并非必不可少。促进LY7吸附需要更高浓度的磷蛋白。LY7细胞对由PRP - 1制备的薄膜的吸附在pH 5至8范围内不受影响。吸附也不受50 mM乳糖的抑制,这与1型菌毛而非2型菌毛起作用的观点一致。黏性放线菌T14、溶牙放线菌ATCC 17982和衣氏放线菌12597也吸附于PRP - 1薄膜,而内氏放线菌ATCC 12104则不吸附。尽管黏性放线菌细胞与吸附的PRP - 1强烈结合,但溶液中PRP - 1或PRP - 3的存在并不抑制黏附。同样地,[³H]PRP - 1不与LY7细胞结合,与该微生物一起孵育时也不被降解。然而,LY7细胞吸附于[³H]PRP - 1薄膜。这些数据表明,当PRP吸附到HA上时,其隐藏的分子片段会暴露;这些片段随后与LY7细胞的黏附素发生反应。黏性放线菌细胞识别仅在表面吸附分子中暴露的PRPs片段的明显能力提供了一种新机制,使该微生物在悬浮于唾液分泌物中时能够附着于牙齿。