4.7 Article

Protein delivery to living cells by thermal stimulation for biophysical investigation

Journal

SCIENTIFIC REPORTS
Volume 12, Issue 1, Pages -

Publisher

NATURE PORTFOLIO
DOI: 10.1038/s41598-022-21103-9

Keywords

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Funding

  1. EC H2020 project iNEXT-Discovery [871037]
  2. Ministero dell'Istruzione, dell'Universita e della Ricerca PRIN [20177XJCHX]
  3. Instruct-ERIC
  4. CERM/CIRMMP Italy

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This article presents a novel protocol that delivers proteins into eukaryotic cells through mild thermal stimulation, allowing for the study of biomolecules in their natural environment. The data demonstrates the effectiveness of this method in delivering proteins to mammalian cells and achieving concentrations suitable for biophysical analysis, such as double electron electron resonance (DEER) measurements for protein conformation characterization.
Studying biomolecules in their native environment represents the ideal sample condition for structural biology investigations. Here we present a novel protocol which allows to delivery proteins into eukaryotic cells through a mild thermal stimulation. The data presented herein show the efficacy of this approach for delivering proteins in the intracellular environment of mammalian cells reaching a concentration range suitable for successfully applying biophysical methods, such as double electron electron resonance (DEER) measurements for characterising protein conformations.

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