Biomedical subjects

Felicidad Almazan

Publications and source records attributed to Felicidad Almazan.

3 recordsLinked to original sources

PubMed2006

The CC chemokine MCP-1 stimulates surface expression of CX3CR1 and enhances the adhesion of monocytes to fractalkine/CX3CL1 via p38 MAPK.

The membrane-anchored form of CX3CL1 has been proposed as a novel adhesion protein for leukocytes. This functional property of CX3CL1 is mediated through CX3CR1, a chemokine receptor expressed predominantly on circulating white blood cells. Thus far, it is still uncertain at what stage of the trafficking process CX3CR1 becomes importantly involved and how the CX3CR1-dependent adhesion of leukocytes is regulated during inflammation. The objective of this study was to examine the functional effects of chemokine stimulation on CX3CR1-mediated adhesion of human monocytes. Consistent with previous reports, our data indicate that the activity of CX3CR1 on resting monocytes is sufficient to mediate cell adhesion to CX3CL1. However, the basal, nonstimulated adhesion activity is low, and we hypothesized that like the integrins, CX3CR1 may require a preceding activation step to trigger firm leukocyte adhesion. Compatible with this hypothesis, stimulation of monocytes with MCP-1 significantly increased their adhesion to immobilized CX3CL1, under both static and physiological flow conditions. The increase of the adhesion activity was mediated through CCR2-dependent signaling and obligatory activation of the p38 MAPK pathway. Stimulation with MCP-1 also induced a rapid increase of CX3CR1 protein on the cell surface. Inhibition of the p38 MAPK pathway prevented this increase of CX3CR1 surface expression and blunted the effect of MCP-1 on cell adhesion, indicating a causal link between receptor surface density and adhesion activity. Together, our data suggest that a chemokine signal is required for firm CX3CR1-dependent adhesion and demonstrate that CCR2 is an important regulator of CX3CL1-dependent leukocyte adhesion.

Animals↗

PubMed2005

The amino terminus and the third extracellular loop of CX3CR1 contain determinants critical for distinct receptor functions.

The G protein-coupled receptor CX3CR1 is a specific receptor for the CX3C chemokine fractalkine (CX3CL1 according to the new chemokine nomenclature). The aim of this study was to identify receptor elements that contribute independently to agonist binding and receptor activation. Targeted mutation of selected acidic amino acid residues demonstrated that the binding activity of CX3CR1 was critically dependent on the two negatively charged residues Asp25 and Glu254 located on the N-terminal domain and third extracellular loop, respectively. In addition, mutation of the uncharged polar residue Tyr14 in the amino terminus caused a reduction in the ligand binding affinity. In contrast, the three acidic residues Glu13, Asp16, and Asp266 did not contribute to ligand binding but were crucial for receptor activation. The mutant receptors E13A, D16A, and D266A bound fractalkine with high affinity but were unable to induce signaling events necessary to support chemotaxis. These acidic residues may engage in electrostatic interactions with basic residues on fractalkine that are necessary for receptor function but not for binding. Our data are consistent with a model of chemokine receptor activation consisting of a multi-step mechanism. Step one mediates the high-affinity fractalkine binding involving Tyr14, Asp25, and Glu254. The initial interaction then triggers the engagement of Glu13, Asp16, and Asp266, which are necessary for CX3CR1 activation.

Amino Acid Sequence↗

PubMed2005

Phosphocholine as a pattern recognition ligand for CD36.

We have previously shown that CD36 recognizes oxidation products of phospholipids on oxidized LDL (OxLDL) such as 1-palmitoyl-2-(5'-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC). The current study was designed to examine whether the phosphocholine (PC) headgroup in POVPC constitutes an obligatory binding target for CD36. To examine the contribution of PC in the binding of POVPC to CD36, we used well-defined synthetic oxidized phospholipids (OxPLs) cross-linked to BSA or to a hexapeptide. The OxPL adducts were then tested for their ability to bind to CD36-transfected cells and for their ability to inhibit OxLDL binding to CD36. Both POVPC-BSA and POVPC-peptide adducts were high-affinity ligands for CD36 and potent inhibitors of OxLDL binding. Enzymatic removal of the entire PC moiety of the POVPC-peptide, or of the choline headgroup alone, as well as substitution of the choline headgroup by ethanolamine abrogated the inhibitory activity of POVPC. Interestingly, PC by itself or cross-linked to BSA did not show any intrinsic competition activity. In conclusion, our data demonstrate that the PC headgroup of OxPL alone is sufficient for binding to CD36, but only if presented in the correct conformation as in OxPL of OxLDL or as in POVPC-peptide adducts.

Animals↗