Temperature-jump microscopy and interaction of Hsp70 heat shock protein with a client protein in vivo

Aniket Ravan, Samuel Procopio, Yann R. Chemla, Martin Gruebele

Research output: Contribution to journalArticlepeer-review

Abstract

Biomolecular processes such as protein–protein interactions can depend strongly on cell type and even vary within a single cell type. Here we develop a microscope with a Peltier-controlled temperature stage, a laser temperature jump to induce heat stress, and an autofocusing feature to mitigate temperature drift during experiments, to study a protein–protein interaction in a selected cell type within a live organism, the zebrafish larva. As an application of the instrument, we show that there is considerable cell-to-cell variation of the heat shock protein Hsp70 binding to one of its clients, phosphoglycerate kinase in vivo. We adapt a key feature from our previous folding study, rare transformation of cells within the larva, so that individual cells can be imaged and differentiated for cell-to-cell response. Our approach can be extended to other organisms and cell types than the ones demonstrated in this work.

Original languageEnglish (US)
Pages (from-to)154-164
Number of pages11
JournalMethods
Volume231
DOIs
StatePublished - Nov 2024

Keywords

  • Fluorescence microscopy
  • Protein–protein interaction
  • Quinary structure
  • Zebrafish

ASJC Scopus subject areas

  • Molecular Biology
  • General Biochemistry, Genetics and Molecular Biology

Fingerprint

Dive into the research topics of 'Temperature-jump microscopy and interaction of Hsp70 heat shock protein with a client protein in vivo'. Together they form a unique fingerprint.

Cite this