先进的RPL19-TRAPKI-seq方法揭示了生物活性化合物的作用机制
Di Zhu1, Junchi Hu2, Renke Tan3
1Laboratory of Tumor Immunology, Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, 200032, China.
Natural products and bioprospecting
|March 5, 2025
概括
一种新的RPL19-TRAPKI-seq方法通过分析翻译mRNA来准确识别药物点. 这种工具准确地揭示了小分子机制,有助于药物发现和开发.
科学领域:
- 分子生物学分子生物学
- 药理学 药理学是指药理学的学科.
- 药物发现 药物发现 药物发现
背景情况:
- 天然产品对于药物开发至关重要,但往往缺乏明确的分子标和研究的作用机制.
- 研究药物机制需要精确的方法来分析分子标和细胞反应.
研究的目的:
- 开发和验证一种新的方法,RPL19-TRAPKI-seq,用于研究小分子作用机制.
- 为了确定可用药物的点,并阐明天然产品类型的抗瘤活性背后的机制.
主要方法:
- 开发了RPL19-TRAPKI-seq,通过结合CRISPR/Cas9敲进和翻译核糖体亲和力净化 (TRAP) 技术.
- 建立了一种表达EGFP-RPL19的稳定细胞系,用于高效的核糖体丰富和对翻译mRNA的分析.
- 使用拉巴胺素 (mTOR抑制剂) 验证了该方法,并研究了抗瘤化合物SBF-1.
主要成果:
- RPL19-TRAPKI-seq精确地丰富了核糖体和翻译mRNA,密切反映了蛋白质表达水平.
- 该方法证实了已知的拉巴胺素作用,并揭示了SBF-1的抗瘤活性源于扰乱细胞氧化酸化.
- 确定了七种适合TRAP的核糖体蛋白质,其中RPL19显示出高的丰富效率.
结论:
- RPL19-TRAPKI-seq是精确阐明小分子作用机制的强大工具.
- 这种方法增强了对小分子的探索,推动了药理学和治疗开发.
- 方便识别天然产品及其类型的分子标.
相关概念视频
Cotranslational Protein Translocation
7.1K
Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
7.1K
LTR Retrotransposons
17.3K
LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
17.3K
Non-LTR Retrotransposons
11.3K
As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
11.3K
Protein Translocation Machinery on the ER Membrane
4.4K
The translocon complex situated on the ER membrane is the main gateway for the protein secretory pathway. It facilitates the transport of nascent peptides into the ER lumen and their insertion into the ER membrane.
Sec61 protein conducting channel
In eukaryotes, the translocon complex comprises a core heterotrimeric translocator channel called the Sec61 complex. This channel includes three transmembrane proteins, Sec61α, Sec61β, and Sec61γ, and is the largest subunit of the...
Sec61 protein conducting channel
In eukaryotes, the translocon complex comprises a core heterotrimeric translocator channel called the Sec61 complex. This channel includes three transmembrane proteins, Sec61α, Sec61β, and Sec61γ, and is the largest subunit of the...
4.4K


