IMMUNOCHEMICAL STUDIES ON BLOOD GROUPS .26. ISOLATION OF OLIGOSACCHARIDES FROM HUMAN OVARIAN CYST BLOOD GROUP A SUBSTANCE INCLUDING 2 DISACCHARIDES AND A TRISACCHARIDE INVOLVED IN SPECIFICITY OF BLOOD GROUP A ANTIGENIC DETERMINANT
IMMUNOCHEMICAL STUDIES ON BLOOD GROUPS .26. ISOLATION OF OLIGOSACCHARIDES FROM HUMAN OVARIAN CYST BLOOD GROUP A SUBSTANCE INCLUDING 2 DISACCHARIDES AND A TRISACCHARIDE INVOLVED IN SPECIFICITY OF BLOOD GROUP A ANTIGENIC DETERMINANT
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DOI：
10.1021/ja00860a018
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发表时间：
1962-01-01
影响因子：
15
通讯作者：
LESKOWITZ, S
中科院分区：
文献类型：
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作者：
SCHIFFMAN, G;KABAT, EA;LESKOWITZ, S
在温和条件下（pH 1.9 - 1.6，100 ℃，2小时），将来自人卵巢囊肿液的A血型物质依次水解。将可透析组分在木炭、纸和玻璃纤维片上分级分离，得到三种二糖和三糖。通过用硼氢化钠还原、高碘酸盐氧化和旋光前后的分析，这些化合物显示为：（1）α-N-乙酰半乳糖氨基酰基-（1- 3）半乳糖，（2）α-半乳糖基-（1 - 3）-N-乙酰葡糖胺，（3）0-半乳糖基-（1- 4）-N-乙酰葡糖胺和（4）α-N-乙酰半乳糖氨基酰基-（1 - 3）-β-半乳糖基-（1- 3）-N-乙酰葡糖胺。该三糖在抑制A-抗-A沉淀方面比以前报道的最有活性的二糖更有活性，可能是A血型抗原决定簇的前三个单元。早期的研究表明，在A血型物质中发现的四种单糖单元中，只有N-乙酰半乳糖胺能够抑制抗-A对A红细胞的血凝作用，2 A物质沉淀抗A抗体8和A血型物质的酶失活4。5从这些观察结果可以推断，N-乙酰半乳糖胺是A特异性寡糖决定簇中的末端非还原单位。α-N-乙酰氨基半乳糖酰-（1-*· 3）-半乳糖的分离（0.1 N HCl 3小时或1 N HCl 0.5小时，100 ℃）的A型血物质6，以及它在比α-甲基-N-乙酰氨基半乳糖低得多的浓度下特异性抑制抗A红细胞的血红蛋白的报告7，强烈表明这种化合物，磅代表从A特异性决定簇的非还原端开始的前两个糖单元。本实验室早期的研究表明，温和的水解条件（在100 ℃下，pH值为1.5至2.0，持续2小时）可将A和B血型物质中的活性基团裂解，留下不可透析的残留物PI。从观察到的PI组分的这些性质可以推断出水解条件的温和性：（1）不可透析性和醇沉淀性，（2）与原始血型物质相比，与马XIV型抗马球菌血清的交叉反应性增加，（3）免疫后在人体内产生循环沉淀素的能力。9温和的
Blood group A substance derived from human ovarian cyst fluid has been sequentially hydrotyzed under mild conditions (pH 1.9 to 1.6 for 2 hr. at 100). The dialyzable components were fractionated on charcoal, paper and glass fiber sheets to yield three disaccharides and a trisaccharide. By analysis before and after reduction with sodium borohydride, periodate oxidation and optical rotation these compounds were shown to be:(1) aN-acetylgalactosaminoyl-(1— 3) galactose,(2) ß-galactosyl-(l—* 3)-N-acetylglucosamine,(3) 0-galactosyl-(1— 4)-N-acetylglucosamine and (4)«-N-acetylgalactosaminoyl-(1-* 3)-/3-galactosyl-(1—* 3)-N-acetylglucosamine. The trisaccharide was shown to be more active in inhibition of A-anti-A precipitation than the most active disaccharide previously reported and is probably the first three units of the blood group A antigenic determinant.Earlier studies have shown that of the four monosaccharide units found in blood group A substance, only N-acetylgalactosamine was capable of inhibiting the hemagglutination of A redcells by anti-A, 2 the precipitation of anti-A by A sub-stance8 and the enzymatic inactivation4 of blood group A substance. 5 From these observations it was inferred that N-acetylgalactosamine was the terminal non-reducing unit in an A specific oligo-saccharide determinant. The isolation of aN-acetylgalactosaminoyl-(l—*· 3)-galactose from a partial acid hydrolysate (0.1 N HC1 for 3 hr. or 1 N HC1 for 0.5 hr. at 100) of blood group A substance6 and the report7 that it specifically inhibited hemag-glutination of A erythrocytes by anti-A at a much lower concentration than does a-methyl-N-acetyl-galactosaminide strongly suggests that this com-pound represents the first two sugar units from the non-reducing end of the A specific determinant. Earlier studies from this Laboratory haveshown that milderhydrolytic conditions (£ H 1.5 to 2.0 for 2 hr. at 100 8) cleave active groupings from blood group A and B substances leaving a non-dialyzable residue, PI. The mildness of the hydrolytic conditions can be inferred from these observed proper-ties of thePI fraction:(1) non-dialyzability and alcohol precipitability,(2) increased cross-reactivity with horse type XIV anti-pneumoccocal sera as compared to the original bloodgroup substance,(3) ability to produce circulating precipitins in humans following immunization. 9 These mild