Membrane Localization of HspA1A, a Stress Inducible 70-kDa Heat-Shock Protein, Depends on Its Interaction with Intracellular Phosphatidylserine

Andrei D. Bilog, Larissa Smulders, Ryan Oliverio, Cedra Labanieh, Julianne Zapanta, Robert Stahelin, Nikolas Nikolaidis

Research output: Contribution to journalArticle

Abstract

HspA1A is a cytosolic molecular chaperone essential for cellular homeostasis. HspA1A also localizes at the plasma membrane (PM) of tumor and stressed cells. However, it is currently unknown how this cytosolic protein translocates to the PM. Taking into account that HspA1A interacts with lipids, including phosphatidylserine (PS), and that lipids recruit proteins to the PM, we hypothesized that the interaction of HspA1A with PS allows the chaperone to localize at the PM. To test this hypothesis, we subjected cells to mild heat-shock and the PM-localized HspA1A was quantified using confocal microscopy and cell surface biotinylation. These experiments revealed that HspA1A's membrane localization increased during recovery from non-apoptotic heat-shock. Next, we selectively reduced PS targets by overexpressing the C2 domain of lactadherin (Lact-C2), a known PS-biosensor, and determined that HspA1A's membrane localization was greatly reduced. In contrast, the reduction of PI(4,5)P2 availability by overexpression of the PLCδ-PH biosensor had minimal effects on HspA1A's PM-localization. Implementation of a fluorescent PS analog, TopFluor-PS, established that PS co-localizes with HspA1A. Collectively, these results reveal that HspA1A's PM localization and anchorage depend on its selective interaction with intracellular PS. This discovery institutes PS as a new and dynamic partner in the cellular stress response.

Original languageEnglish (US)
JournalBiomolecules
Volume9
Issue number4
DOIs
StatePublished - Apr 17 2019
Externally publishedYes

Fingerprint

HSP70 Heat-Shock Proteins
Phosphatidylserines
Cell membranes
Membranes
Cell Membrane
Biosensing Techniques
Biosensors
Shock
Hot Temperature
Biotinylation
Lipids
Molecular Chaperones
Confocal microscopy
Programmable logic controllers
Confocal Microscopy
Tumors
Proteins
Homeostasis
Availability
Recovery

Keywords

  • heat-shock proteins
  • lipid–protein interactions
  • phosphatidylserine
  • plasma membrane

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Membrane Localization of HspA1A, a Stress Inducible 70-kDa Heat-Shock Protein, Depends on Its Interaction with Intracellular Phosphatidylserine. / Bilog, Andrei D.; Smulders, Larissa; Oliverio, Ryan; Labanieh, Cedra; Zapanta, Julianne; Stahelin, Robert; Nikolaidis, Nikolas.

In: Biomolecules, Vol. 9, No. 4, 17.04.2019.

Research output: Contribution to journalArticle

Bilog, Andrei D. ; Smulders, Larissa ; Oliverio, Ryan ; Labanieh, Cedra ; Zapanta, Julianne ; Stahelin, Robert ; Nikolaidis, Nikolas. / Membrane Localization of HspA1A, a Stress Inducible 70-kDa Heat-Shock Protein, Depends on Its Interaction with Intracellular Phosphatidylserine. In: Biomolecules. 2019 ; Vol. 9, No. 4.
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