Tronchin G, Bouchara J P, Robert R, Senet J M
Laboratoire d'Immunologie-Parasitologie, UER des Sciences Médicales et Pharmaceutiques, Angers, France.
Infect Immun. 1988 Aug;56(8):1987-93. doi: 10.1128/iai.56.8.1987-1993.1988.
Germ tubes of Candida albicans produced an additional fibrillar surface layer responsible for enhanced adherence to plastic. The correlation between germination of C. albicans and adherence of germ tubes to a plastic matrix led us to consider the existence of germ tube-specific adhesive components involved in the attachment process. Using concanavalin A-sensitized latex microspheres, we first detected extracellular molecules on the plastic surface after removal of the adherent germ tubes. Electron microscopy confirmed that fibrils of the germ tube involved in cell-substratum interconnections were retained on the plastic surface. Cytochemistry with concanavalin A-gold labeling demonstrated that these fibrillar structures contained mannoproteins. Dithiothreitol and iodoacetamide treatment of washed plastic allowed us to further characterize these fibrillar adhesins. Through analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two components with molecular weights (MWs) of 68,000 and 60,000 were detected on the plastic surface. The 68,000-MW component appeared to be one of the major constituents of the germ tube surface layers. Biosynthetic labeling experiments performed with L-[35S]methionine revealed two additional proteins: a high-MW component (greater than 200,000), and a 200,000-MW component. These four proteins, strongly labeled on the plastic surface and on the germ tube cell wall layers, were in contrast slightly labeled or even nonidentified in the culture supernatant, suggesting their involvement in germ tube adherence.
白色念珠菌的芽管产生了一层额外的纤维状表面层,这层表面层增强了其对塑料的黏附。白色念珠菌的萌发与芽管对塑料基质的黏附之间的相关性,使我们考虑在附着过程中存在芽管特异性黏附成分。我们首先使用伴刀豆球蛋白A致敏的乳胶微球,在去除黏附的芽管后检测塑料表面的细胞外分子。电子显微镜证实,参与细胞与基质连接的芽管纤维保留在了塑料表面。伴刀豆球蛋白A-金标记的细胞化学实验表明,这些纤维状结构含有甘露糖蛋白。用二硫苏糖醇和碘乙酰胺处理洗涤过的塑料,使我们能够进一步表征这些纤维状黏附素。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析,在塑料表面检测到两种分子量分别为68,000和60,000的成分。分子量为68,000的成分似乎是芽管表面层的主要成分之一。用L-[35S]甲硫氨酸进行的生物合成标记实验揭示了另外两种蛋白质:一种高分子量成分(大于200,000)和一种分子量为200,000的成分。这四种蛋白质在塑料表面和芽管细胞壁层上被强烈标记,相比之下,在培养上清液中则被轻微标记甚至未被识别,这表明它们参与了芽管的黏附。