Partial characterization of the sexual agglutination factor from Hansenula wingei Y-2340 type 5 cells.
Partial characterization of the sexual agglutination factor from Hansenula wingei Y-2340 type 5 cells.
复制标题
温氏汉逊酵母 Y-2340 5 型细胞性凝集因子的部分表征。
DOI：
10.1021/bi00708a030
复制
发表时间：
1974
期刊：
影响因子：
2.9
通讯作者：
C. Ballou
中科院分区：
文献类型：
--
作者：
P. Yen;C. Ballou
摘要：采用亲和层析和凝胶过滤的方法纯化了温氏汉氏菌Y-2340 5型细胞上的性凝集因子（5-agglutinin），该因子经枯草菌素消化从细胞表面释放出来。该制剂的分子量为9.6 X 105，由85%的碳水化合物（主要是甘露糖），10%的蛋白质和5%的磷酸盐组成。蛋白质部分含有55%的丝氨酸和6-8%的苏氨酸，其中85%的这两种氨基酸在5-凝集素用0.1 n NaOH在23小时处理24小时时被破坏，这种条件促进了取代的丝氨酸和苏氨酸单位的消除。在此处理过程中，附着在凝集素上的90%的碳水化合物被释放为甘露寡糖，具有1-15个糖单位，主要片段是八糖。因此，5-凝集素是一种新型糖蛋白，其中60%的氨基酸被碳水化合物取代，其结构与H. wingei [P.]的总细胞壁甘露聚糖蛋白非常不同。H. Yen和CE Ballou(1974)，生物化学13,2420)，其中大部分碳水化合物以长支链的形式存在于天冬酰胺上。尽管存在这些差异，5-凝集素和全细胞壁甘露聚糖在其乙酰分解模式、甲基化分析、高酸盐消耗和免疫学研究中给出了相似的结果，所有这些都表明碳水化合物片段中的键和构型非常相似。5-Agglutinin是
Pauline Hsiao Yen and Clinton E. Ballou* abstract: The sexual agglutination factor on Hansenula wingei Y-2340 type 5 cells (5-agglutinin) was purified by affinity chromatography and gel filtration after its release from the cell surface by subtilisin digestion. The preparation had a molecular weight of 9.6 X 105, and it was composed of 85% carbohydrate (mostly mannose), 10% protein, and 5% phosphate. The protein part contained 55% serine and 6-8% threonine, and 85% of these two amino acids was destroyed on treatment of 5-agglutinin with 0.1 n NaOH at 23 for 24 hr, conditions that promote ß elimination of substituted serine and threonine units. During this treatment, 90% of the carbohydrate attached to the agglutinin was released as mannooligosaccharides with 1-15 sugar units, the principal fragment being the octasaccharide. Thus, 5-agglutinin is a novel glycoprotein in which 60% of the amino acids are sub-stituted by carbohydrate, a structure very different from that of the total cell wall mannan-protein of H. wingei [P. H. Yen and CE Ballou (1974), Biochemistry 13, 2420] in which most of the carbohydrate is present as long branched poly-saccharide chains attached to asparagine. In spite of these differences, the 5-agglutinin and the whole cell wall mannan gave similar results in their acetolysis patterns, methylation analysis, periodate consumption, and immunological studies, all suggesting that the linkages and configurations in the carbohydrate fragments were very similar. 5-Agglutinin was