Syntaxin 11 regulates the stimulus-dependent transport of Toll-like receptor 4 to the plasma membrane by cooperating with SNAP-23 in macrophages

Molecular Biology of the Cell
Daiki KinoshitaKiyotaka Hatsuzawa

Abstract

Syntaxin 11 (stx11) is a soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) that is selectively expressed in immune cells; however, its precise role in macrophages is unclear. We showed that stx11 knockdown reduces the phagocytosis of Escherichia coli in interferon-γ-activated macrophages. stx11 knockdown decreased Toll-like receptor 4 (TLR4) localization on the plasma membrane without affecting total expression. Plasma membrane-localized TLR4 was primarily endocytosed within 1 h by lipopolysaccharide (LPS) stimulation and gradually relocalized 4 h after removal of LPS. This relocalization was significantly impaired by stx11 knockdown. The lack of TLR4 transport to the plasma membrane is presumably related to TLR4 degradation in acidic endosomal organelles. Additionally, an immunoprecipitation experiment suggested that stx11 interacts with SNAP-23, a plasma membrane-localized SNARE protein, whose depletion also inhibits TLR4 replenishment in LPS-stimulated cells. Using an intramolecular Förster resonance energy transfer (FRET) probe for SNAP-23, we showed that the high FRET efficiency caused by LPS stimulation is reduced by stx11 knockdown. These findings suggest that stx11 regulates the stimulus-depe...Continue Reading

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Citations

Sep 23, 2020·Journal of Cellular Physiology·Bor L Tang
Sep 5, 2020·Yonago Acta Medica·Kiyotaka Hatsuzawa, Chiye Sakurai
Oct 16, 2020·Cellular and Molecular Life Sciences : CMLS·Anna CiesielskaKatarzyna Kwiatkowska

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Datasets Mentioned

BETA
GM130

Methods Mentioned

BETA
GTPase
transfection
Fluorescence
FRET
reverse transcription PCR
PCR

Software Mentioned

GraphPad Prism
ImageJ
GraphPad

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