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相关概念视频

Initiation of Translation02:33

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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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从不同的人类细胞类型中进行高效的无细胞翻译.

Jana Ziegelmüller1, Nikolaos Kouvelas2, Nino Schwaller1

  • 1Department of Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, Bern, Switzerland.

The Journal of biological chemistry
|May 30, 2025
PubMed
概括

研究人员开发了一种可扩展的方法,用于使用双离心法进行无细胞翻译系统. 这种技术可以从各种人体细胞系中获得高质量的溶解物,从而更好地研究蛋白质合成调节.

关键词:
没有细胞的翻译.细胞类型特定的溶解物.通过双离心法进行离心.在体外翻译 in vitro翻译蛋白质合成 蛋白质合成合成生物学 合成生物学翻译法规的翻译法规

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科学领域:

  • 分子生物学分子生物学
  • 生物化学 生物化学

背景情况:

  • 无细胞翻译系统对于研究蛋白质合成和翻译调节至关重要.
  • 目前从多种细胞类型中制备无细胞系统的局限性阻碍了对上下文依赖的监管机制的研究.

研究的目的:

  • 开发一种可扩展的方法,从各种人类细胞系中生成对翻译有能力的细胞溶解物.
  • 为了使细胞特异性转化调节在不同的生物环境中的研究.

主要方法:

  • 采用双离心技术来制备细胞溶解物.
  • 溶解条件优化了附着细胞和悬浮细胞培养物.
  • 酸盐是由HEK-293 (粘合剂和悬浮液),HeLa,SH-SY5Y和U2OS细胞系制备的.

主要成果:

  • 从多个人类细胞系成功产生了高质量的,对翻译有能力的溶解体.
  • 发现细胞特异性因素影响翻译效率,附着HeLa细胞表现出最高的活性.
  • 不同的细胞系对溶解条件的敏感性各不相同,突出显示了对参数优化的需要.

结论:

  • 开发的双离心法为产生细胞类型特异性溶解物提供了强大的和可适应的方法.
  • 这一进步扩大了体外翻译系统的实用性,用于研究复杂的翻译机制.
  • 这些发现强调了细胞特异性优化对于无细胞系统中高效蛋白质生产的重要性.