CO-TRANSLATIONAL FOLDING OF NASCENT PROTEINS

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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.