Manning M, Mitchell T G
Infect Immun. 1980 Nov;30(2):484-95. doi: 10.1128/iai.30.2.484-495.1980.
The extent of the macromolecular change accompanying yeast to mycelium morphogenesis of Candida albicans was analyzed by two-dimensional gel electrophoresis of the cytoplasmic proteins of the two growth forms after antibody cross-absorption experiments. Pure cultures of yeasts and true hyphae (i.e., without concomitant production of pseudohyphae) were grown in a synthetic low-sulfate medium (LSM). The two strains selected for this study were strain 4918, which produces pure mycelial (M) cultures in LSM at 37 degrees C (designated 4918-37M) and yeasts (Y) at 24 degrees C (4918-24Y), and strain 2252, which produces yeasts exclusively at both 24 and 37 degrees C in LSM (2252-24Y and 2252-37Y). The proteins of both strains were labeled at both temperatures with [35S]sulfate, and cytoplasmic fractions were prepared by mechanical disruption and ultracentrifugation. Rabbits were immunized with the 4918-24Y and 4918-37M cytoplasmic fractions to produce anti-yeast-phase and anti-mycelial-phase hyperimmune sera. Each radiolabeled cytoplasmic fraction was absorbed with anti-mycelial-phase immunoglobulin, anti-yeast immunoglobulin, and immunoglobulin from normal rabbit serum. Staphyloccal protein A was used to remove immune complexes. The labeled, nonabsorbed proteins were also analyzed by two-dimensional electrophoresis. Highly reproducible protein spot patterns were obtained which defined hundreds of proteins in each extract. The specificity of the immunoglobulin hundreds of proteins in each extract. The specificity of the immunoglobulin preparations was extremely broad, and as many as 168 cytoplasmic antigens were detected. Eighty-three antigens were recognized in the mycelial-phase extract only by the anti-mycelial-phase immunoglobulin. However, comparative analysis revealed that all of these proteins were present in at least one other extract. Therefore, none of them was unique to the mycelial morphology. Eleven antigens were detected in the 2252-37Y extract that were not present in the extracts from strain 4918, which indicates that proteins obtained from different strains may express similar antigenic determinants, but differ in their physiochemical properties.
通过抗体交叉吸收实验后对两种生长形式的细胞质蛋白进行二维凝胶电泳，分析了白色念珠菌从酵母形态转变为菌丝形态过程中伴随的大分子变化。酵母和真正菌丝（即不伴有假菌丝产生）的纯培养物在合成低硫酸盐培养基（LSM）中生长。本研究选用的两个菌株分别是菌株4918，其在37℃的LSM中产生纯菌丝体（M）培养物（命名为4918 - 37M），在24℃产生酵母（Y）培养物（4918 - 24Y）；以及菌株2252，其在LSM中于24℃和37℃均只产生酵母（2252 - 24Y和2252 - 37Y）。两个菌株的蛋白质在这两个温度下均用[35S]硫酸盐进行标记，并通过机械破碎和超速离心制备细胞质组分。用4918 - 24Y和4918 - 37M细胞质组分免疫兔子，以产生抗酵母期和抗菌丝期的超免疫血清。每个放射性标记的细胞质组分用抗菌丝期免疫球蛋白、抗酵母免疫球蛋白和正常兔血清中的免疫球蛋白进行吸收。使用葡萄球菌蛋白A去除免疫复合物。对标记的、未吸收的蛋白质也进行二维电泳分析。获得了高度可重复的蛋白质斑点图谱，每个提取物中可鉴定出数百种蛋白质。免疫球蛋白制剂的特异性非常广泛，检测到多达168种细胞质抗原。只有抗菌丝期免疫球蛋白能在菌丝期提取物中识别出83种抗原。然而，对比分析表明所有这些蛋白质至少在其他一种提取物中也存在。因此，它们中没有一种是菌丝形态所特有的。在2252 - 37Y提取物中检测到11种在菌株4918的提取物中不存在的抗原，这表明从不同菌株获得的蛋白质可能表达相似的抗原决定簇，但在其理化性质上有所不同。
