Biochemical characterization of the interaction between HspA1A and phospholipids

Cell Stress & Chaperones
Chelsea McCallisterNikolas Nikolaidis

Abstract

Seventy-kilodalton heat shock proteins (Hsp70s) are molecular chaperones essential for maintaining cellular homeostasis. Apart from their indispensable roles in protein homeostasis, specific Hsp70s localize at the plasma membrane and bind to specific lipids. The interaction of Hsp70s with lipids has direct physiological outcomes including lysosomal rescue, microautophagy, and promotion of cell apoptosis. Despite these essential functions, the Hsp70-lipid interactions remain largely uncharacterized. In this study, we characterized the interaction of HspA1A, an inducible Hsp70, with five phospholipids. We first used high concentrations of potassium and established that HspA1A embeds in membranes when bound to all anionic lipids tested. Furthermore, we found that protein insertion is enhanced by increasing the saturation level of the lipids. Next, we determined that the nucleotide-binding domain (NBD) of the protein binds to lipids quantitatively more than the substrate-binding domain (SBD). However, for all lipids tested, the full-length protein is necessary for embedding. We also used calcium and reaction buffers equilibrated at different pH values and determined that electrostatic interactions alone may not fully explain the as...Continue Reading

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Citations

Mar 1, 2016·Biochemical and Biophysical Research Communications·Chelsea McCallisterNikolas Nikolaidis
Apr 15, 2016·Cell Stress & Chaperones·Victor LopezAntonio De Maio
Aug 20, 2020·Physical Chemistry Chemical Physics : PCCP·Madhumitha DhanasekaranSoumit S Mandal
Jan 8, 2020·International Journal of Environmental Research and Public Health·Beatriz MatosMário S Diniz
Aug 23, 2020·International Journal of Molecular Sciences·Larissa SmuldersNikolas Nikolaidis
Nov 6, 2018·Biochemical and Biophysical Research Communications·Ryan OliverioNikolas Nikolaidis
Sep 4, 2021·Cell Stress & Chaperones·Antonio De Maio, Lawrence Hightower

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