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Super-Resolution Spatial Proximity Detection with Proximity-PAINT.

ABSTRACT: Visualizing the functional interactions of biomolecules such as proteins and nucleic acids is key to understanding cellular life on the molecular scale. Spatial proximity is often used as a proxy for the direct interaction of biomolecules. However, current techniques to visualize spatial proximity are either limited by spatial resolution, dynamic range, or lack of single-molecule sensitivity. Here, we introduce Proximity-PAINT (pPAINT), a variation of the super-resolution microscopy technique DNA-PAINT. pPAINT uses a split-docking-site configuration to detect spatial proximity with high sensitivity, low false-positive rates, and tunable detection distances. We benchmark and optimize pPAINT using designer DNA nanostructures and demonstrate its cellular applicability by visualizing the spatial proximity of alpha- and beta-tubulin in microtubules using super-resolution detection.

SUBMITTER: Schueder F

PROVIDER: S-EPMC7839522 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Super-Resolution Spatial Proximity Detection with Proximity-PAINT.

Schueder Florian F Lara-Gutiérrez Juanita J Haas Daniel D Beckwith Kai Sandvold KS Yin Peng P Ellenberg Jan J Jungmann Ralf R

Angewandte Chemie (International ed. in English) 20201109 2

Visualizing the functional interactions of biomolecules such as proteins and nucleic acids is key to understanding cellular life on the molecular scale. Spatial proximity is often used as a proxy for the direct interaction of biomolecules. However, current techniques to visualize spatial proximity are either limited by spatial resolution, dynamic range, or lack of single-molecule sensitivity. Here, we introduce Proximity-PAINT (pPAINT), a variation of the super-resolution microscopy technique DN ...[more]

PMID: 32936507

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