title: Dynamic regulation of CD45 lateral mobility by the spectrin-ankyrin cytoskeleton of T cells.
abstract: The leukocyte common antigen, CD45, is a critical immune regulator whose activity is modulated by cytoskeletal interactions. Components of the spectrin-ankyrin cytoskeleton have been implicated in the trafficking and signaling of CD45. We have examined the lateral mobility of CD45 in resting and activated T lymphocytes using single-particle tracking and found that the receptor has decreased mobility caused by increased cytoskeletal contacts in activated cells. Experiments with cells that have disrupted betaI spectrin interactions show decreased cytoskeletal contacts in resting cells and attenuation of receptor immobilization in activated cells. Applying two types of population analyses to single-particle tracking trajectories, we find good agreement between the diffusion coefficients obtained using either a mean squared displacement analysis or a hidden Markov model analysis. Hidden Markov model analysis also reveals the rate of association and dissociation of CD45-cytoskeleton contacts, demonstrating the importance of this analysis for measuring cytoskeleton binding events in live cells. Our findings are consistent with a model in which multiple cytoskeletal contacts, including those with spectrin and ankyrin, participate in the regulation of CD45 lateral mobility. These interactions are a major factor in CD45 immobilization in activated cells. Furthermore, cellular activation leads to CD45 immobilization by reduction of the CD45-cytoskeleton dissociation rate. Short peptides that mimic spectrin repeat domains alter the association rate of CD45 to the cytoskeleton and cause an apparent decrease in dissociation rates. We propose a model for CD45-cytoskeleton interactions and conclude that the spectrin-ankyrin-actin network is an essential determinant of immunoreceptor mobility.
authors.affiliation: Department of Chemistry and Alberta Ingenuity Centre for Carbohydrate Science, University of Alberta, Edmonton, Alberta T6G 2G2, Canada. ccairo@ualberta.ca
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Cairo
authors.forename: Raibatak
authors.initials: R
authors.lastname: Das
authors.forename: Amgad
authors.initials: A
authors.lastname: Albohy
authors.forename: Quentin J
authors.initials: QJ
authors.lastname: Baca
authors.forename: Deepti
authors.initials: D
authors.lastname: Pradhan
authors.forename: Jon S
authors.initials: JS
authors.lastname: Morrow
authors.forename: Daniel
authors.initials: D
authors.lastname: Coombs
authors.forename: David E
authors.initials: DE
authors.lastname: Golan
mesh_terms: D017487:Ankyrins / Q000737:chemistry*; D003599:Cytoskeleton / Q000378:metabolism*; D004058:Diffusion; D006801:Humans; D019169:Jurkat Cells; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D008390:Markov Chains; D008856:Microscopy, Fluorescence / Q000379:methods; D008863:Microspheres; D008954:Models, Biological; D008958:Models, Molecular; D008968:Molecular Conformation; D017434:Protein Structure, Tertiary; D013049:Spectrin / Q000737:chemistry*; D013601:T-Lymphocytes / Q000378:metabolism*
chemical_list: D017487:Ankyrins; D013049:Spectrin; D017493:Leukocyte Common Antigens