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用DDD-固蓝B染色法对艾氏腹水癌细胞进行蛋白质巯基和活性二硫键的定量细胞光谱测定(作者译)

[Quantitative cytospectrometrical determination of protein sulfhydryl groups and reactive disulfide groups by the DDD-Fast blue B-staining method on Ehrlich ascites tumor cells (author's transl)].

作者信息

Nöhammer G

出版信息

Acta Histochem. 1977;59(2):317-28.

PMID:73341
Abstract

Protein sulfhydryl groups are histochemically demonstrated by reacting with DDD followed by coupling with Fast blue B. The molar absorptivity of the formed azo dye is 19000 per mole SH reacted. DDD simultaneously reacts with protein-SH- and -SS-groups. However, the reaction with SH-groups is approximately 1000 times faster than that observed with SS-groups. With Ehrlich ascites tumour cells the reaction of DDD with SH-groups is completed within 7 h while the reaction of DDD with SS-groups needs 14 days for completion. Due to the extreme difference in the reaction rates protein bound SH-groups as well as reactive SS-groups can be estimated quantitatively by cytospectrophotometrical methods. The cells investigated showed an average SH-content of (1,30 +/- 0,03) X 10(-14) M SH/cell while the average content of reactive SS-groups was (1,59 +/- 0,28) X 10(-14) M SS/cell. In addition it was found that especially the amount of reactive SS-groups per cell is not constant but exhibits seasonal variations.

摘要

蛋白质巯基通过与DDD反应,然后与固蓝B偶联进行组织化学显示。所形成的偶氮染料的摩尔吸光系数为每摩尔反应的巯基19000。DDD能同时与蛋白质的巯基和二硫键基团反应。然而,与巯基的反应比与二硫键基团的反应快约1000倍。对于艾氏腹水瘤细胞,DDD与巯基的反应在7小时内完成,而DDD与二硫键基团的反应则需要14天才能完成。由于反应速率存在极大差异,结合蛋白质的巯基以及活性二硫键基团均可通过细胞分光光度法进行定量测定。所研究的细胞显示,平均巯基含量为(1.30±0.03)×10⁻¹⁴ M SH/细胞,而活性二硫键基团的平均含量为(1.59±0.28)×10⁻¹⁴ M SS/细胞。此外,还发现尤其每个细胞中活性二硫键基团的数量并非恒定不变,而是呈现季节性变化。

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