Regulated shedding of transmembrane chemokines by the disintegrin and metalloproteinase 10 facilitates detachment of adherent leukocytes
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Regulated shedding of transmembrane chemokines by the disintegrin and metalloproteinase 10 facilitates detachment of adherent leukocytes. / Hundhausen, Christian; Schulte, Alexander; Schulz, Beate; Andrzejewski, Michael G; Schwarz, Nicole; von Hundelshausen, Philipp; Winter, Ulrike; Paliga, Krzysztof; Reiss, Karina; Saftig, Paul; Weber, Christian; Ludwig, Andreas.
In: J IMMUNOL, Vol. 178, No. 12, 15.06.2007, p. 8064-72.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - Regulated shedding of transmembrane chemokines by the disintegrin and metalloproteinase 10 facilitates detachment of adherent leukocytes
AU - Hundhausen, Christian
AU - Schulte, Alexander
AU - Schulz, Beate
AU - Andrzejewski, Michael G
AU - Schwarz, Nicole
AU - von Hundelshausen, Philipp
AU - Winter, Ulrike
AU - Paliga, Krzysztof
AU - Reiss, Karina
AU - Saftig, Paul
AU - Weber, Christian
AU - Ludwig, Andreas
PY - 2007/6/15
Y1 - 2007/6/15
N2 - CX3CL1 (fractalkine) and CXCL16 are unique members of the chemokine family because they occur not only as soluble, but also as membrane-bound molecules. Expressed as type I transmembrane proteins, the ectodomain of both chemokines can be proteolytically cleaved from the cell surface, a process known as shedding. Our previous studies showed that the disintegrin and metalloproteinase 10 (ADAM10) mediates the largest proportion of constitutive CX3CL1 and CXCL16 shedding, but is not involved in the phorbolester-induced release of the soluble chemokines (inducible shedding). In this study, we introduce the calcium-ionophore ionomycin as a novel, very rapid, and efficient inducer of CX3CL1 and CXCL16 shedding. By transfection in COS-7 cells and ADAM10-deficient murine embryonic fibroblasts combined with the use of selective metalloproteinase inhibitors, we demonstrate that the inducible generation of soluble forms of these chemokines is dependent on ADAM10 activity. Analysis of the C-terminal cleavage fragments remaining in the cell membrane reveals multiple cleavage sites used by ADAM10, one of which is preferentially used upon stimulation with ionomycin. In adhesion studies with CX3CL1-expressing ECV-304 cells and cytokine-stimulated endothelial cells, we demonstrate that induced CX3CL1 shedding leads to the release of bound monocytic cell lines and PBMC from their cellular substrate. These data provide evidence for an inducible release mechanism via ADAM10 potentially important for leukocyte diapedesis.
AB - CX3CL1 (fractalkine) and CXCL16 are unique members of the chemokine family because they occur not only as soluble, but also as membrane-bound molecules. Expressed as type I transmembrane proteins, the ectodomain of both chemokines can be proteolytically cleaved from the cell surface, a process known as shedding. Our previous studies showed that the disintegrin and metalloproteinase 10 (ADAM10) mediates the largest proportion of constitutive CX3CL1 and CXCL16 shedding, but is not involved in the phorbolester-induced release of the soluble chemokines (inducible shedding). In this study, we introduce the calcium-ionophore ionomycin as a novel, very rapid, and efficient inducer of CX3CL1 and CXCL16 shedding. By transfection in COS-7 cells and ADAM10-deficient murine embryonic fibroblasts combined with the use of selective metalloproteinase inhibitors, we demonstrate that the inducible generation of soluble forms of these chemokines is dependent on ADAM10 activity. Analysis of the C-terminal cleavage fragments remaining in the cell membrane reveals multiple cleavage sites used by ADAM10, one of which is preferentially used upon stimulation with ionomycin. In adhesion studies with CX3CL1-expressing ECV-304 cells and cytokine-stimulated endothelial cells, we demonstrate that induced CX3CL1 shedding leads to the release of bound monocytic cell lines and PBMC from their cellular substrate. These data provide evidence for an inducible release mechanism via ADAM10 potentially important for leukocyte diapedesis.
KW - ADAM Proteins
KW - Amyloid Precursor Protein Secretases
KW - Animals
KW - COS Cells
KW - Cell Adhesion
KW - Cell Membrane
KW - Cercopithecus aethiops
KW - Chemokine CX3CL1
KW - Chemokines, CX3C
KW - Chemokines, CXC
KW - Disintegrins
KW - Humans
KW - Leukocytes
KW - Matrix Metalloproteinase 10
KW - Membrane Proteins
KW - Mice
KW - Receptors, Scavenger
KW - Transfection
M3 - SCORING: Journal article
C2 - 17548644
VL - 178
SP - 8064
EP - 8072
JO - J IMMUNOL
JF - J IMMUNOL
SN - 0022-1767
IS - 12
ER -