Yamada H, Aramaki Y, Miyazaki T
Biochim Biophys Acta. 1977 Apr 27;497(2):396-407. doi: 10.1016/0304-4165(77)90197-0.
A non-dyalyzable specific agglutination factor of myxamoebae obtained from culture broth during the growth phase of Dictyostelium discoideum NC-4 was thermostable but the agglutination activity disappeared below pH 5.0. In the case of formalinized myxamoebae, digestion of the factor with Pronase decreased the activity, but periodate treatment of the factor did not affect the activity. Myxamoebal agglutination by this factor was inhibited by the addition of uronic acid, polyuronide (protuberic acid), and cell-surface polysaccharide prepared from the myxamoebae, but the agglutination was not affected by citric acid or glycine. The factor was purified by ethanol precipitation, column chromatography using DEAE-cellulose and Sepharose-2B, and zone electrophoresis. Chemical analysis of the purified factor gave 61.0% carbohydrate and 26.1% protein, and glucose, mannose, xylose and rhamnose (molar ratios of 9,3 : 3.2 : 2.1 : 1.0) were detected as the component sugars. The content of uronic acid was 12.9%. When the myxamoebae of the growth phase were starved in Millipore-supporting medium, the agglutination activity was detected in the supernatant of the medium.
在盘基网柄菌NC - 4生长阶段从培养液中获得的一种不可透析的黏液变形虫特异性凝集因子具有热稳定性，但在pH 5.0以下凝集活性消失。对于经甲醛固定的黏液变形虫，用链霉蛋白酶消化该因子会降低活性，但用高碘酸盐处理该因子不影响活性。添加糖醛酸、聚糖醛酸（原结核菌素酸）和从黏液变形虫制备的细胞表面多糖可抑制该因子引起的黏液变形虫凝集，但柠檬酸或甘氨酸不影响凝集。该因子通过乙醇沉淀、使用DEAE - 纤维素和琼脂糖 - 2B的柱色谱法以及区带电泳进行纯化。对纯化因子的化学分析表明，其碳水化合物含量为61.0%，蛋白质含量为26.1%，检测到葡萄糖、甘露糖、木糖和鼠李糖（摩尔比为9.3 : 3.2 : 2.1 : 1.0）作为组成糖类。糖醛酸含量为12.9%。当处于生长阶段的黏液变形虫在微孔支持培养基中饥饿时，在培养基的上清液中检测到凝集活性。
