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海胆卵和胚胎中的磷酸酶活性抗原。II. 未受精卵与长腕幼虫活性的比较。

Phosphatase active antigens in sea urchin eggs and embryos. II. A comparison between the activities in unfertilized eggs and plutei.

作者信息

Westin M

出版信息

J Exp Zool. 1975 Jun;192(3):315-22. doi: 10.1002/jez.1401920304.

Abstract

Phosphatase activities in sea urchin eggs and plutei were investigated by means of histochemical staining of immunoprecipitates. Two protein fractions were obtained by extraction in a hypotonic medium and by detergent treatment of the residual pellet. Three distinctly different phosphatase activities were discerned, nucleoside diphosphatase (EC 3.6.1.6.), acid phosphatase (EC 3.1.3.2.) and alkaline phosphatase (EC 3.1.3.1.). The nucleoside diphosphatase activity, which was confined to one antigen, was present in both water soluble and detergent extracts and at roughly the same concentration in eggs and plutei. By means of a monospecific antiserum the immunological identify of this antigen was established in all instances. The acid phosphatase activity, which was displayed by ten detergent extracted antigens in eggs, was only found in five detergent extracted antigens in plutei. This decrease in number of enzyme active antigens was also reflected by a general decrease in number of enzyme active antigens was also reflected by a general decrease in activity as assessed by quantitative determinations. Furthermore, by means of absorbed antisera it was established that two or three of the acid phosphatase active antigens were "egg specific". Another acid phosphatase active antigen, which was common to both developmental stages, was investigated by a monospecific antiserum. While this antigen was found in both soluble fractions, it was only enzymatically active when extracted with detergent. Alkaline phosphatase active antigens were only found in the detergent extract of plutei. However, immunoprecipitates with this activity appeared both with antiserum against unfertilized eggs and with antiserum against plutei. This suggests that the egg contained the antigens in an enzymatically inactive form.

摘要

通过免疫沉淀物的组织化学染色研究了海胆卵和幼体中的磷酸酶活性。通过在低渗培养基中提取以及对残留沉淀进行去污剂处理,获得了两种蛋白质组分。辨别出三种明显不同的磷酸酶活性,即核苷二磷酸酶(EC 3.6.1.6.)、酸性磷酸酶(EC 3.1.3.2.)和碱性磷酸酶(EC 3.1.3.1.)。局限于一种抗原的核苷二磷酸酶活性存在于水溶性提取物和去污剂提取物中,在卵和幼体中的浓度大致相同。通过单特异性抗血清在所有情况下都确定了该抗原的免疫学特性。卵中由十种去污剂提取的抗原所显示的酸性磷酸酶活性,在幼体中仅在五种去污剂提取的抗原中发现。酶活性抗原数量的这种减少也反映在通过定量测定评估的活性普遍下降上。此外,通过吸收抗血清确定,酸性磷酸酶活性抗原中的两到三种是“卵特异性的”。另一种在两个发育阶段都共有的酸性磷酸酶活性抗原,用单特异性抗血清进行了研究。虽然在两种可溶性组分中都发现了这种抗原,但只有在用去污剂提取时才具有酶活性。碱性磷酸酶活性抗原仅在幼体的去污剂提取物中发现。然而,具有这种活性的免疫沉淀物在用未受精卵的抗血清和幼体的抗血清时都出现了。这表明卵中含有无酶活性形式的抗原。

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