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Updated: Jun 1, 2025

Measurement of Chitinase Activity in Biological Samples

Measurement of Chitinase Activity in Biological Samples

Published on: August 22, 2019

表达和功能分析的小鼠基因酶没有ZZ域的黄金葡萄球菌蛋白A蛋白

Masahiro Kimura¹, Takumi Sakoh², Masayoshi Sakaguchi²

¹Division of Infectious Diseases and Immunology, Department of Microbiology, School of Medicine, Iwate Medical University, 1-1-1 Idaidori, Yahaba, Iwate 028-3694, Japan; Department of Chemistry and Life Science, Kogakuin University, 2665‑1 Nakano‑Cho, Hachioji, Tokyo 192-0015, Japan.

International journal of biological macromolecules

|January 17, 2025

概括

研究人员开发了没有ZZ域的新版本的小鼠基因酶 (Chit1和AMCase),以改善免疫研究. 这些功能性重组基因酶是研究哺乳动物免疫力和真菌相互作用的宝贵工具.

科学领域:

生物化学生物化学
免疫学免疫学免疫学
分子生物学分子生物学

背景情况:

酸酶对于哺乳动物的免疫反应至关重要,特别是在分解真菌细胞壁时.
一些基因酶中的ZZ域可以通过与抗体的Fc区域结合来干扰免疫学试验.

研究的目的:

为了表达和表征重组小鼠基托酶 (Chit1) 和缺乏ZZ域的酸性哺乳动物基酶 (AMCase).
评估这些改性基因酶的功能活性和稳定性.
确定它们作为免疫学分析的改进工具的实用性.

主要方法:

在大肠杆菌中,ZZ域被删除的小鼠Chit1和AMCase (Chit1-V5-His和AMCase-V5-His) 的重组表达.
使用合成和天然基质进行基因溶解活性测试.
不同扫描度和热稳定性测试.
对Fc受体介导的细胞分裂的干扰的评估.

主要成果:

在没有ZZ域的情况下成功表达了可溶性,功能性重组基因酶 (Chit1-V5-His和AMCase-V5-His).
奇特1-V5-His与其含有ZZ域的对应物相比,具有可比的奇丁溶解活性和功能稳定性.
AMCase-V5-His在pH值为2.0时表现出最佳的活性,与天然的AMCase相一致.
删除ZZ域的基因酶并没有阻碍真菌的Fc受体介导的细胞分裂.

结论:

开发的表达系统有效地产生了缺少ZZ域的功能性小鼠基因酶.
由于没有Fc结合干扰,这些修饰的基提纳酶是免疫学研究的优越工具.
这有助于进一步研究基因酶在哺乳动物免疫力和宿主-病原体相互作用中的作用.

关键词:

基因酸酶的使用方法热稳定性热稳定性 ZZ 域名 ZZ 域名

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