CO-TRANSLATIONAL FOLDING OF NASCENT PROTEINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 本项目的目标是用核磁共振技术探索核糖体结合的新生多肽的构象和动力学。这种类型的研究将为所讨论的新生蛋白质的共翻译折叠机制提供实验证据。核糖体的核磁共振研究迄今很少，因为它的旋转相关时间较长，导致谱线展宽。然而，对于核糖体结合的新生多肽链来说，这个问题不那么严格，这些多肽链延伸到远离核糖体的地方，预计本质上是高度动态的。通过在无细胞系统中创建新生的链-核糖体复合体，提供所需的15N和13C标记的氨基酸，选择性地对新生链进行同位素富集化。该项目利用NMRFAM高场磁铁和低温探头资源，这些资源非常适合解决相关的敏感性问题。这项研究的重点是从抹香鲸去肌红蛋白中提取的核糖体结合的新生链条。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of this project is to explore the conformation and dynamics of ribosome-bound nascent polypeptides by Nuclear Magnetic Resonance (NMR). This type of study will generate experimental evidence for the co-translational folding mechanism of the nascent protein in question. NMR studies on the ribosome have been scarce so far due to its long rotational correlation time and the resulting spectral line broadening. This issue, however, is less stringent for ribosome-bound nascent polypeptide chains, which extend away from the ribosome and are expected to be highly dynamic in nature. Nascent chains are selectively isotopically enriched by creating nascent chain-ribosome complexes in cell free systems, supplied with the desired 15N and 13C-labeled amino acids. This project makes use of the NMRFAM high field magnet and cryoprobe resources, which are ideally suited to address the relevant sensitivity issues. The research focuses on ribosome-bound nascent chains derived from sperm whale apomyoglobin.