CO-TRANSLATIONAL FOLDING OF NASCENT PROTEINS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本研究的目的是利用核磁共振技术研究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由于其长的旋转相关时间和由此产生的谱线展宽而至今很少。然而，这个问题对于核糖体结合的新生多肽链不太严格，其延伸远离核糖体并且预期在自然界中是高度动态的。新生链通过在无细胞系统中产生新生链-核糖体复合物而被选择性地同位素富集，所述无细胞系统被供应有所需的15 N和13 C标记的氨基酸。该项目利用了NMRFAM的高场磁体和冷冻探针资源，这些资源非常适合解决相关的敏感性问题。这项研究的重点是来自抹香鲸脱辅基肌红蛋白的核糖体结合新生链。

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.