Every protein inside a cell must fold into a precise three-dimensional shape to work, and this folding process is riddled with errors that can lead to disease. Current lab methods study proteins in isolation, missing the crowded, chaperone-filled environment where folding actually happens. This project will map the real routes proteins take as they fold inside living cells—using stalled ribosomes to catch proteins mid-folding, then tracking which chaperones they meet and how the ribosome itself guides the process. If successful, this work will bridge a fundamental gap between test-tube biochemistry and real biology. Understanding how cells manage protein folding could eventually explain why it fails in conditions like Alzheimer’s, Parkinson’s, or cystic fibrosis, where misfolded proteins accumulate. It may also reveal why some proteins fold efficiently while others need help, informing future drug design or synthetic biology efforts. This is primarily fundamental science. The immediate payoff is a clearer picture of a core cellular process, not a product or therapy. But past discoveries in protein folding—like the role of heat-shock proteins—have already shaped cancer treatments and neurodegenerative disease research. This project lays the groundwork for similar long-term advances.
View original technical description
Proteins are synthesized on ribosomes as linear chains of amino acids that in most cases fold into unique 3-D structures to function. The folding process is intrinsically error-prone, and how it is accomplished efficiently represents a problem of fundamental biological and medical importance. Protein folding is highly regulated in vivo. Folding and oligomeric assembly are tightly coupled to synthesis, and occur in the context of an extensive network of molecular chaperones - proteins that help other proteins fold. Structurally and functionally diverse chaperones cooperate to form a robust network that coordinates with the ribosome to promote protein folding, reverse misfolding, and inhibit toxic aggregation. Belying this complexity, most current approaches to resolve folding pathways are limited to studying single proteins in isolation, and therefore fail to recapitulate the rich landscape of protein maturation in vivo. The key outstanding questions are: i) what routes do different proteins take through the chaperone network? And ii) how does the organization of the network, coupled to the process of translation, influence folding pathways? I aim to define and resolve authentic protein biogenesis pathways using structural proteomics. We will: 1. Use stalled translation intermediates to identify sequential chaperone interactions for a set of cytosolic proteins in E.coli and human cells. 2. Reconstitute coupled translation/folding reactions in vitro, and use these to quantitatively describe the contribution of different biogenesis factors to protein folding. 3. Use state-of-the-art hydrogen/deuterium exchange- and chemical crosslinking-mass spectrometry to precisely define the folding pathways of nascent proteins on the ribosome, and in complex with chaperones. I anticipate that these results will bridge the gap between in vitro and in vivo concepts of protein folding, and contribute to a new understanding of how the cellular environment informs protein biogenesis.
Plain English summaries and category classifications on this site are generated by AI and may not perfectly reflect the original research.
Is something wrong? Let us know