Phosphatidic acid induces conformational changes in Sec18 protomers that prevent SNARE priming

Matthew L. Starr, Robert P. Sparks, Andres S. Arango, Logan R. Hurst, Zhiyu Zhao, Muyun Lihan, Jermaine L. Jenkins, Emad Tajkhorshid, Rutilio Fratti

Research output: Contribution to journalArticle

Abstract

Eukaryotic cell homeostasis requires transfer of cellular components among organelles and relies on membrane fusion catalyzed by SNARE proteins. Inactive SNARE bundles are reactivated by hexameric N-ethylmaleimide-sensitive factor, vesicle-fusing ATPase (Sec18/NSF)-driven disassembly that enables a new round of membrane fusion. We previously found that phosphatidic acid (PA) binds Sec18 and thereby sequesters it from SNAREs and that PA dephosphorylation dissociates Sec18 from the membrane, allowing it to engage SNARE complexes. We now report that PA also induces conformational changes in Sec18 protomers and that hexameric Sec18 cannot bind PA membranes. Molecular dynamics (MD) analyses revealed that the D1 and D2 domains of Sec18 contain PA-binding sites and that the residues needed for PA binding are masked in hexameric Sec18. Importantly, these simulations also disclosed that a major conformational change occurs in the linker region between the D1 and D2 domains, which is distinct from the conformational changes that occur in hexameric Sec18 during SNARE priming. Together, these findings indicate that PA regulates Sec18 function by altering its architecture and stabilizing membrane-bound Sec18 protomers.

Original languageEnglish (US)
Pages (from-to)3100-3116
Number of pages17
JournalJournal of Biological Chemistry
Volume294
Issue number9
DOIs
StatePublished - Jan 1 2019

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SNARE Proteins
Phosphatidic Acids
Protein Subunits
Membranes
N-Ethylmaleimide-Sensitive Proteins
Membrane Fusion
Fusion reactions
Eukaryotic Cells
Molecular Dynamics Simulation
Organelles
Molecular dynamics
Homeostasis
Binding Sites

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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Phosphatidic acid induces conformational changes in Sec18 protomers that prevent SNARE priming. / Starr, Matthew L.; Sparks, Robert P.; Arango, Andres S.; Hurst, Logan R.; Zhao, Zhiyu; Lihan, Muyun; Jenkins, Jermaine L.; Tajkhorshid, Emad; Fratti, Rutilio.

In: Journal of Biological Chemistry, Vol. 294, No. 9, 01.01.2019, p. 3100-3116.

Research output: Contribution to journalArticle

Starr, Matthew L. ; Sparks, Robert P. ; Arango, Andres S. ; Hurst, Logan R. ; Zhao, Zhiyu ; Lihan, Muyun ; Jenkins, Jermaine L. ; Tajkhorshid, Emad ; Fratti, Rutilio. / Phosphatidic acid induces conformational changes in Sec18 protomers that prevent SNARE priming. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 9. pp. 3100-3116.
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