Splitting of human thyroglobulin. III. Comparison of fragments obtained during enzymatic digestion and by reduction and alkylation.
Splitting of human thyroglobulin. III. Comparison of fragments obtained during enzymatic digestion and by reduction and alkylation.
复制标题
人类甲状腺球蛋白的分裂。
DOI：
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发表时间：
1974
影响因子：
4.6
通讯作者：
N. Rose
中科院分区：
文献类型：
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作者：
P. Mehta;N. Rose
纯化的甲状腺球蛋白用胰蛋白酶和木瓜蛋白酶消化，用二硫苏糖醇还原，用碘乙酰胺烷基化。用免疫学技术对所得片段进行分离和鉴定。酶降解后，分离出一个小片段，称为分数2。其沉降系数约为3S，在SDS存在下，聚丙烯酰胺凝胶电泳测定的摩尔重量约为37,000道尔顿。在Ouchterlony试验中，当兔抗人甲状腺球蛋白时，与完整的甲状腺球蛋白相比，它是抗原缺陷的。与人自身抗血清，分数2未显示任何沉淀反应在Ouchterlony试验。但对晒黑细胞血凝试验有较弱的抑制作用。胰蛋白酶和木瓜蛋白酶消化所得的部分在免疫学上似乎是相同的。然而，当这些部分与还原和烷基化甲状腺球蛋白获得的类似产品相比，后者在Ouchterlony试验中表现出非同构反应，并且在鞣制细胞血凝试验中具有较弱的抑制能力。还原和烷基化所得馏分的沉降系数为8S，摩尔重量约为38,500。可以得出结论，酶分解和还原烷基化得到的小摩尔重量分数是不同的，尽管两者都具有许多被兔抗血清识别的抗原决定因子，而缺乏大多数自身抗原决定因子。
Purified thyroglobulin was digested with trypsin and papain and was also reduced with dithiothreitol and alkylated with iodoacetamide. The resulting fragments were separated and characterized by immunological techniques. Following enzymatic degradation a small fragment, termed fraction 2, was isolated. It had a sedimentation coefficient of approximately 3S and a mol. wt determined by polyacrylamide gel electrophoresis in presence of SDS was approximately 37,000 daltons. In the Ouchterlony test, it was antigenically deficient as compared with intact thyroglobulin, when rabbit antisera to human thyroglobulin were used. With human autoantisera, fraction 2 did not show any precipitin reaction in the Ouchterlony test. However, it produced weak inhibition in the tanned cell haemagglutination test. The fractions obtained from trypsin and papain digestion appeared to be immunologically identical. However, when these fractions were compared with the similar product obtained from reduced and alkylated thyroglobulin, the latter fraction showed a reaction of non-identity in the Ouchterlony test, and had a weaker inhibiting capacity in tanned cell haemagglutination test. The fraction produced by reduction and alkylation had a sedimentation coefficient of 8S and an approximate mol. wt of 38,500. It can be concluded that the small mol. wt fractions derived from enzymatic breakdown and by reduction and alkylation are different although both possess a number of antigenic determinants recognized by the rabbit antiserum and lack most of the autoantigenic determinants.