Structural and biological interaction of HSC-70 protein with phosphatidylserine in endosomal microautophagy

Kateryna Morozova, Cristina C. Clement, Susmita Kaushik, Barbara Stiller, Esperanza Arias-Perez, Atta Ahmad, Jennifer N. Rauch, Victor Chatterjee, Chiara Melis, Brian Scharf, Jason E. Gestwicki, Ana Maria Cuervo, Erik R P Zuiderweg, Laura Santambrogio

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

hsc-70 (HSPA8) is a cytosolic molecular chaperone, which plays a central role in cellular proteostasis, including quality control during protein refolding and regulation of protein degradation. hsc-70 is pivotal to the process of macroautophagy, chaperone-mediated autophagy, and endosomal microautophagy. The latter requires hsc-70 interaction with negatively charged phosphatidylserine (PS) at the endosomal limiting membrane. Herein, by combining plasmon resonance, NMR spectroscopy, and amino acid mutagenesis, we mapped the C terminus of the hsc-70 LID domain as the structural interface interacting with endosomal PS, and we estimated an hsc-70/PS equilibrium dissociation constant of 4.7 ± 0.1 μm. This interaction is specific and involves a total of 4-5 lysine residues. Plasmon resonance and NMR results were further experimentally validated by hsc-70 endosomal binding experiments and endosomal microautophagy assays. The discovery of this previously unknown contact surface for hsc-70 in this work elucidates the mechanism of hsc-70 PS/membrane interaction for cytosolic cargo internalization into endosomes.

Original languageEnglish (US)
Pages (from-to)18096-18106
Number of pages11
JournalJournal of Biological Chemistry
Volume291
Issue number35
DOIs
StatePublished - Aug 26 2016

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Phosphatidylserines
Autophagy
Proteins
Protein Refolding
Membranes
Mutagenesis
Molecular Chaperones
Endosomes
Quality Control
Nuclear magnetic resonance spectroscopy
Proteolysis
Lysine
Quality control
Assays
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Amino Acids
Degradation
Experiments

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)
  • Molecular Biology
  • Cell Biology

Cite this

Structural and biological interaction of HSC-70 protein with phosphatidylserine in endosomal microautophagy. / Morozova, Kateryna; Clement, Cristina C.; Kaushik, Susmita; Stiller, Barbara; Arias-Perez, Esperanza; Ahmad, Atta; Rauch, Jennifer N.; Chatterjee, Victor; Melis, Chiara; Scharf, Brian; Gestwicki, Jason E.; Cuervo, Ana Maria; Zuiderweg, Erik R P; Santambrogio, Laura.

In: Journal of Biological Chemistry, Vol. 291, No. 35, 26.08.2016, p. 18096-18106.

Research output: Contribution to journalArticle

Morozova, K, Clement, CC, Kaushik, S, Stiller, B, Arias-Perez, E, Ahmad, A, Rauch, JN, Chatterjee, V, Melis, C, Scharf, B, Gestwicki, JE, Cuervo, AM, Zuiderweg, ERP & Santambrogio, L 2016, 'Structural and biological interaction of HSC-70 protein with phosphatidylserine in endosomal microautophagy', Journal of Biological Chemistry, vol. 291, no. 35, pp. 18096-18106. https://doi.org/10.1074/jbc.M116.736744
Morozova, Kateryna ; Clement, Cristina C. ; Kaushik, Susmita ; Stiller, Barbara ; Arias-Perez, Esperanza ; Ahmad, Atta ; Rauch, Jennifer N. ; Chatterjee, Victor ; Melis, Chiara ; Scharf, Brian ; Gestwicki, Jason E. ; Cuervo, Ana Maria ; Zuiderweg, Erik R P ; Santambrogio, Laura. / Structural and biological interaction of HSC-70 protein with phosphatidylserine in endosomal microautophagy. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 35. pp. 18096-18106.
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