{"department":[{"_id":"PaSc"}],"type":"book_chapter","day":"13","language":[{"iso":"eng"}],"year":"2024","author":[{"full_name":"Guillerm, Undina","first_name":"Undina","id":"bb74f472-ae54-11eb-9835-bc9c22fb1183","last_name":"Guillerm"},{"last_name":"Sučec","full_name":"Sučec, Iva","first_name":"Iva"},{"id":"7B541462-FAF6-11E9-A490-E8DFE5697425","first_name":"Paul","full_name":"Schanda, Paul","last_name":"Schanda","orcid":"0000-0002-9350-7606"}],"publication_identifier":{"issn":["0076-6879"]},"date_updated":"2024-10-07T08:02:22Z","date_published":"2024-09-13T00:00:00Z","publication_status":"inpress","citation":{"ieee":"U. Guillerm, I. Sučec, and P. Schanda, “Generation of TIM chaperone substrate complexes,” in <i>Methods in Enzymology</i>, Elsevier.","apa":"Guillerm, U., Sučec, I., &#38; Schanda, P. (n.d.). Generation of TIM chaperone substrate complexes. In <i>Methods in Enzymology</i>. Elsevier. <a href=\"https://doi.org/10.1016/bs.mie.2024.07.051\">https://doi.org/10.1016/bs.mie.2024.07.051</a>","mla":"Guillerm, Undina, et al. “Generation of TIM Chaperone Substrate Complexes.” <i>Methods in Enzymology</i>, Elsevier, doi:<a href=\"https://doi.org/10.1016/bs.mie.2024.07.051\">10.1016/bs.mie.2024.07.051</a>.","ama":"Guillerm U, Sučec I, Schanda P. Generation of TIM chaperone substrate complexes. In: <i>Methods in Enzymology</i>. Elsevier. doi:<a href=\"https://doi.org/10.1016/bs.mie.2024.07.051\">10.1016/bs.mie.2024.07.051</a>","short":"U. Guillerm, I. Sučec, P. Schanda, in:, Methods in Enzymology, Elsevier, n.d.","chicago":"Guillerm, Undina, Iva Sučec, and Paul Schanda. “Generation of TIM Chaperone Substrate Complexes.” In <i>Methods in Enzymology</i>. Elsevier, n.d. <a href=\"https://doi.org/10.1016/bs.mie.2024.07.051\">https://doi.org/10.1016/bs.mie.2024.07.051</a>.","ista":"Guillerm U, Sučec I, Schanda P.Generation of TIM chaperone substrate complexes. In: Methods in Enzymology. ."},"article_processing_charge":"No","scopus_import":"1","doi":"10.1016/bs.mie.2024.07.051","oa_version":"None","status":"public","corr_author":"1","_id":"18167","title":"Generation of TIM chaperone substrate complexes","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","date_created":"2024-10-01T10:58:27Z","abstract":[{"text":"Holdase chaperones are essential in the mitochondrial membrane-protein biogenesis as they stabilize preproteins and keep them in an import-competent state as they travel through the aqueous cytosol and intermembrane space. The small TIM chaperones of the mitochondrial intermembrane space function within a fine balance of client promiscuity and high affinity binding, while being also able to release their client proteins without significant energy barrier to the downstream insertases/translocases. The tendency of the preproteins to aggregate and the dynamic nature of the preprotein—chaperone complexes makes the preparation of these complexes challenging. Here we present two optimized methods for complex formation of highly hydrophobic precursor proteins and chaperones: a pull-down approach and an in-vitro translation strategy. In the former, attaching the client protein to an affinity resin keeps the individual client protein copies apart from each other and decreases the client self-aggregation probability, thereby favouring complex formation. In the latter approach, a purified chaperone, added to the cell-free protein synthesis, captures the nascent precursor protein. The choice of method will depend on the desired client-chaperone complex amount, or the need for specific labeling scheme.","lang":"eng"}],"publisher":"Elsevier","quality_controlled":"1","month":"09","publication":"Methods in Enzymology"}