Imaging Proteins Sensitive to Direct Fusions Using Transient Peptide–Peptide Interactions

Zoe Gidden, Curran Oi, Emily j. Johnston, Zuzanna Konieczna, Haresh Bhaskar, Lorena Mendive-Tapia, Fabio De moliner, Susan j. Rosser, Simon g. j. Mochrie, Marc Vendrell, Mathew h. Horrocks, Lynne Regan

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Fluorescence microscopy enables specific visualization of proteins in living cells and has played an important role in our understanding of the protein subcellular location and function. Some proteins, however, show altered localization or function when labeled using direct fusions to fluorescent proteins, making them
difficult to study in live cells. Additionally, the resolution of fluorescence microscopy is limited to ∼200 nm, which is 2 orders of magnitude larger than the size of most proteins. To circumvent these challenges, we previously developed LIVE-PAINT, a live-cell superresolution approach that takes advantage of short interacting peptides to transiently bind a fluorescent protein to the protein-ofinterest. Here, we successfully use LIVE-PAINT to image yeast
membrane proteins that do not tolerate the direct fusion of a fluorescent protein by using peptide tags as short as 5-residues. We also demonstrate that it is possible to resolve multiple proteins at the nanoscale concurrently using orthogonal peptide interaction pairs.
KEYWORDS: membrane protein, protein−protein interaction, super-resolution microscopy, live-cell imaging, LIVE-PAINT, yeast
Original languageEnglish
Pages (from-to)10633-10641
JournalNano Letters
Volume23
Issue number22
Early online date2 Nov 2023
DOIs
Publication statusPublished - 22 Nov 2023

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