不同PIP动机的PCNA分子识别:Tyr211酸化的作用
Antonio Ruiz-Albor1, Belén Chaves-Arquero1, Inés Martín-Barros2
1Centro de Investigaciones Biológicas Margarita Salas (CIB), CSIC, Madrid 28040, Spain.
International journal of biological macromolecules
|June 16, 2024
概括
在Y211的增殖细胞核抗原 (PCNA) 酸化对关键调节蛋白p21和p15的结合影响最小. 这种修改也影响与DNA聚合酶 δ (p125) 和USP29的结合,但亲和力非常低.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 细胞生物学 细胞生物学
背景情况:
- 增殖细胞核抗原 (PCNA) 对于真核细胞DNA复制和修复至关重要,协调酶和调控蛋白质.
- PCNA的功能是通过翻译后的修改来调节的,包括在Y211的酸化,这会影响DNA损伤.
- 许多PCNA相互作用蛋白通过保存的PCNA相互作用蛋白 (PIP) 基因结合.
研究的目的:
- 调查Y211酸化对PCNA识别p21和p15中的正规PIP动机的影响.
- 描述PCNA与DNA聚合酶 δ (p125) 和USP29.29的非正规PIP序列的结合.
- 了解Y211酸化如何影响PCNA-蛋白相互作用的亲和力和特异性.
主要方法:
- 准备PCNA与p-carboxymethyl-L-phenylalanine (pCMF) 在Y211以模仿酸化.
- 生物化学试验测量PCNA与p21,p15,p125和USP29.29.2的结合亲和力.
- 使用生物物理技术分析PCNA-PIP动机相互作用.
主要成果:
- Y211酸化对PCNA对p21 (nM亲和度) 和p15 (μM亲和度) 的正规PIP序列的分子识别和结合亲和度的影响最小.
- 对p125和USP29的非正规PIP序列的PCNA结合发生在与正规PIP相同的位置.
- 发现PCNA对p125和USP29的PIP序列的结合亲和力非常低.
结论:
- Y211对PCNA的酸化不会显著改变其与正规PIP含蛋白p21和p15的相互作用.
- 虽然p125和USP29的非正规PIP序列与PCNA结合,但它们的亲和力比正规PIPs要低得多.
- 该研究提供了通过翻译后修改和各种蛋白质相互作用对PCNA功能的细微调节的见解.
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