At the immunological synapse, the activated leukocyte cell adhesion molecule (ALCAM) on a dendritic cell (DC) and CD6 molecules on a T cell contribute to sustained DC-T-cell contacts. However, little is known about how ALCAM-CD6 bonds resist and adapt to mechanical stress. Here, we combine single-cell force spectroscopy (SCFS) with total-internal reflection fluorescence microscopy to examine ALCAM-CD6-mediated cell adhesion. The combination of cells expressing ALCAM constructs with certain cytoplasmic tail mutations and improved SCFS analysis processes reveal that the affinity of ALCAM-CD6 bonds is not influenced by the linking of the intracellular domains of ALCAM to the actin cortex. By contrast, the recruitment of ALCAM to adhesion sites and the propensity of ALCAM to anchor plasma membrane tethers depend on actin cytoskeletal interactions. Furthermore, linking ALCAM to the actin cortex through adaptor proteins stiffens the cortex and strengthens cell adhesion. We propose a framework for how ALCAMs contribute to DC-T-cell adhesion, stabilize DC-T-cell contacts and form a mechanical link between CD6 and the actin cortex to strengthen cell adhesion at the immunological synapse.
Cloning, mapping, and characterization of activated leukocyte-cell adhesion molecule (ALCAM), a CD6 ligand
Ezrin/radixin/moesin (ERM) proteins bind to a positively charged amino acid cluster in the juxta-membrane cytoplasmic domain of CD44, CD43, and ICAM-2
Dynamic regulation of activated leukocyte cell adhesion molecule-mediated homotypic cell adhesion through the actin cytoskeleton
Subsecond induction of alpha4 integrin clustering by immobilized chemokines stimulates leukocyte tethering and rolling on endothelial vascular cell adhesion molecule 1 under flow conditions
Molecular basis for the homophilic activated leukocyte cell adhesion molecule (ALCAM)-ALCAM interaction
Ezrin is a downstream effector of trafficking PKC-integrin complexes involved in the control of cell motility
Dynamic interaction of VCAM-1 and ICAM-1 with moesin and ezrin in a novel endothelial docking structure for adherent leukocytes
Cytoskeletal restraints regulate homotypic ALCAM-mediated adhesion through PKCalpha independently of Rho-like GTPases
Measurement of the anchorage force between GPI-anchored alkaline phosphatase and supported membranes by AFM force spectroscopy
Lymph node chemokines promote sustained T lymphocyte motility without triggering stable integrin adhesiveness in the absence of shear forces
Distinct kinetic and mechanical properties govern ALCAM-mediated interactions as shown by single-molecule force spectroscopy
A bond for a lifetime: employing membrane nanotubes from living cells to determine receptor-ligand kinetics
Hotspots of GPI-anchored proteins and integrin nanoclusters function as nucleation sites for cell adhesion.
Association of syntenin-1 with M-RIP polarizes Rac-1 activation during chemotaxis and immune interactions.
United we stand: integrating the actin cytoskeleton and cell-matrix adhesions in cellular mechanotransduction
Investigating differential cell-matrix adhesion by directly comparative single-cell force spectroscopy
Assay for characterizing the recovery of vertebrate cells for adhesion measurements by single-cell force spectroscopy
Synchronizing atomic force microscopy force mode and fluorescence microscopy in real time for immune cell stimulation and activation studies
Kindlin-2 cooperates with talin to activate integrins and induces cell spreading by directly binding paxillin
AFM force spectroscopy reveals how subtle structural differences affect the interaction strength between Candida albicans and DC-SIGN
Mechanical Stimulation of Piezo1 Receptors Depends on Extracellular Matrix Proteins and Directionality of Force
Activated leukocyte cell adhesion molecule (ALCAM) is a marker of recurrence and promotes cell migration, invasion, and metastasis in early-stage endometrioid endometrial cancer
Proteome analysis of human CD56neg NK cells reveals a homogeneous phenotype surprisingly similar to CD56dim NK cells
Human T-Lymphotropic Virus Type 1-Induced Overexpression of Activated Leukocyte Cell Adhesion Molecule (ALCAM) Facilitates Trafficking of Infected Lymphocytes through the Blood-Brain Barrier
Alternative splicing of ALCAM enables tunable regulation of cell-cell adhesion through differential proteolysis
Increasing throughput of AFM-based single cell adhesion measurements through multisubstrate surfaces
Syntenin-1 and ezrin proteins link activated leukocyte cell adhesion molecule to the actin cytoskeleton.
ALCAM shedding at the invasive front of the tumor is a marker of myometrial infiltration and promotes invasion in endometrioid endometrial cancer
N-glycan mediated adhesion strengthening during pathogen-receptor binding revealed by cell-cell force spectroscopy
Two CD9 tetraspanin family members of Japanese flounder (Paralichthys olivaceus): characterization and comparative analysis of the anti-infectious immune function.
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