Event Detail (Archived)
Driving Biological Discovery: An Expanding Toolkit for Affinity Proteomics
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John LaCava, Ph.D., research assistant professor, The Rockefeller University
- Speaker bio(s)
In this talk, John LaCava will briefly highlight some recent work on the proteomic and biochemical characterization of human LINE-1 retrotransposons in model cells, as well as ongoing attempts to characterize LINE-1 ribonucleoproteins immunoprecipitated from resected colorectal cancers. Increasingly, he aims to bring basic research tools to bear on clinical samples. Can studies of the LINE-1 vicinal interactome inform hypotheses concerning the mechanics of its pathobiological effects in tumor?
Yet, objectives extend beyond tabulating lists of candidate "smoking guns.” One long term objective is to transfer cognate healthy and pathogenic macromolecules from their in vivo milieus into test tubes for further downstream study. While there are many methods that can detect (directly or indirectly) the existence of protein interactions, there are few options that permit investigators to retrieve and transfer intact physiological interactions from the cell into an appropriately stabilizing in vitro environment. The distinction is significant: in the latter case, the parts of the cell are physically isolated and enriched in a form that can be subjected to further fruitful downstream research, including biochemical, enzymological, and structural study. Retrieving complexes from the native source is often nontrivial but has some very important practical benefits: it avoids (i) the requirement of advanced knowledge of their complete composition and (ii) artifacts associated with recombinant expression and exogenous reconstitution. Unfortunately, there is no way to predict, a priori, the best way to transfer any given macromolecule from its natural milieu into a suitable in vitro environment. To address this challenge, LaCava and his colleagues have developed a screen for interactome mining, based on affinity proteomics, that is akin to a crystallographic screen; and the platform is under continuous development. He will summarize the screening approach, including its latest improvements and future directions, leveraging a couple research vignettes to do so.
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- Lelia Saba
- (212) 327-8136