Cytoskeletal confinement of CX3CL1 limits its susceptibility to proteolytic cleavage by ADAM10

CX3CL1 is a unique chemokine that acts both as a transmembrane endothelial adhesion molecule and, upon proteolytic cleavage, a soluble chemoattractant for circulating leukocytes. The constitutive release of soluble CX3CL1 requires the interaction of its transmembrane species with the integral membra...

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Bibliographic Details
Published in:Molecular biology of the cell 2014-12, Vol.25 (24), p.3884-3899
Main Authors: Wong, Harikesh S, Jaumouillé, Valentin, Heit, Bryan, Doodnauth, Sasha A, Patel, Sajedabanu, Huang, Yi-Wei, Grinstein, Sergio, Robinson, Lisa A
Format: Article
Language:English
Subjects:
Actin Cytoskeleton - metabolism
ADAM Proteins - metabolism
ADAM10 Protein
Amyloid Precursor Protein Secretases - metabolism
Cell Membrane - metabolism
Cells, Cultured
Chemokine CX3CL1 - genetics
Chemokine CX3CL1 - metabolism
Endocytosis - drug effects
Human Umbilical Vein Endothelial Cells - metabolism
Humans
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Membrane Proteins - metabolism
Microscopy, Fluorescence
Protein Binding
Proteolysis
Tumor Necrosis Factor-alpha - pharmacology
Videotape Recording
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Summary:CX3CL1 is a unique chemokine that acts both as a transmembrane endothelial adhesion molecule and, upon proteolytic cleavage, a soluble chemoattractant for circulating leukocytes. The constitutive release of soluble CX3CL1 requires the interaction of its transmembrane species with the integral membrane metalloprotease ADAM10, yet the mechanisms governing this process remain elusive. Using single-particle tracking and subdiffraction imaging, we studied how ADAM10 interacts with CX3CL1. We observed that the majority of cell surface CX3CL1 diffused within restricted confinement regions structured by the cortical actin cytoskeleton. These confinement regions sequestered CX3CL1 from ADAM10, precluding their association. Disruption of the actin cytoskeleton reduced CX3CL1 confinement and increased CX3CL1-ADAM10 interactions, promoting the release of soluble chemokine. Our results demonstrate a novel role for the cytoskeleton in limiting membrane protein proteolysis, thereby regulating both cell surface levels and the release of soluble ligand.
ISSN:1059-1524
1939-4586
DOI:10.1091/mbc.E13-11-0633