Department of Clinical Chemistry, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
School of Biomedical Engineering, Faculty of Engineering and Information Technology, University of Technology Sydney, Sydney, Australia.
Sci Data. 2021 Apr 12;8(1):100. doi: 10.1038/s41597-021-00884-0.
Progesterone receptor (PR) isoforms, PRA and PRB, act in a progesterone-independent and dependent manner to differentially modulate the biology of breast cancer cells. Here we show that the differences in PRA and PRB structure facilitate the binding of common and distinct protein interacting partners affecting the downstream signaling events of each PR-isoform. Tet-inducible HA-tagged PRA or HA-tagged PRB constructs were expressed in T47DC42 (PR/ER negative) breast cancer cells. Affinity purification coupled with stable isotope labeling of amino acids in cell culture (SILAC) mass spectrometry technique was performed to comprehensively study PRA and PRB interacting partners in both unliganded and liganded conditions. To validate our findings, we applied both forward and reverse SILAC conditions to effectively minimize experimental errors. These datasets will be useful in investigating PRA- and PRB-specific molecular mechanisms and as a database for subsequent experiments to identify novel PRA and PRB interacting proteins that differentially mediated different biological functions in breast cancer.
孕激素受体（PR）异构体 PRA 和 PRB 以孕激素非依赖性和依赖性方式发挥作用，从而差异化调节乳腺癌细胞的生物学功能。在这里，我们展示了 PRA 和 PRB 结构的差异促进了常见和独特的蛋白相互作用伙伴的结合，影响每个 PR-异构体的下游信号事件。在 T47DC42（PR/ER 阴性）乳腺癌细胞中表达 tet 诱导的 HA 标记的 PRA 或 HA 标记的 PRB 构建体。亲和纯化与稳定同位素标记的氨基酸在细胞培养中的技术（SILAC）质谱技术相结合，全面研究了非配体和配体条件下 PRA 和 PRB 的相互作用伙伴。为了验证我们的发现，我们应用了正向和反向 SILAC 条件，有效地最小化了实验误差。这些数据集将有助于研究 PRA 和 PRB 特异性的分子机制，并作为随后的实验数据库，以识别新的 PRA 和 PRB 相互作用蛋白，这些蛋白在乳腺癌中差异介导不同的生物学功能。
