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

doi:10.1021/acs.nanolett.3c03780

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 journal › Article › peer-review

Abstract

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 language	English
Pages (from-to)	10633-10641
Journal	Nano Letters
Volume	23
Issue number	22
Early online date	2 Nov 2023
DOIs	https://doi.org/10.1021/acs.nanolett.3c03780
Publication status	Published - 22 Nov 2023

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10.1021/acs.nanolett.3c03780

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Final published version, 4.23 MBLicence: Creative Commons: Attribution (CC-BY)

https://pubs.acs.org/doi/10.1021/acs.nanolett.3c03780

Dynamic Activatable Fluorophores
Vendrell Escobar, M. (Principal Investigator)
EU government bodies
1/06/18 → 31/05/23
Project: Research

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@article{b8d9c71e611b4af08a24aa73c33312bb,

title = "Imaging Proteins Sensitive to Direct Fusions Using Transient Peptide–Peptide Interactions",

abstract = "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",

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

year = "2023",

month = nov,

day = "22",

doi = "10.1021/acs.nanolett.3c03780",

language = "English",

volume = "23",

pages = "10633--10641",

journal = "Nano Letters",

issn = "1530-6984",

publisher = "American Chemical Society",

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T1 - Imaging Proteins Sensitive to Direct Fusions Using Transient Peptide–Peptide Interactions

AU - Gidden, Zoe

AU - Oi, Curran

AU - Johnston, Emily j.

AU - Konieczna, Zuzanna

AU - Bhaskar, Haresh

AU - Mendive-Tapia, Lorena

AU - De moliner, Fabio

AU - Rosser, Susan j.

AU - Mochrie, Simon g. j.

AU - Vendrell, Marc

AU - Horrocks, Mathew h.

AU - Regan, Lynne

PY - 2023/11/22

Y1 - 2023/11/22

N2 - 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

AB - 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

U2 - 10.1021/acs.nanolett.3c03780

DO - 10.1021/acs.nanolett.3c03780

M3 - Article

SN - 1530-6984

VL - 23

SP - 10633

EP - 10641

JO - Nano Letters

JF - Nano Letters

IS - 22

ER -

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

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Dynamic Activatable Fluorophores

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