Concanavalin A induces a cytoskeletal association of T200 molecules in T lymphocytes

A recent report indicated that T200 molecules interact with elements of the cytoskeleton in BW5147 T lymphoma cells. We have confirmed the cytoskeletal association of T200 by examining nonionic detergent-soluble and detergent-insoluble fractions of murine T cell tumor cell lines, cloned cytotoxic T lymphocyte lines, and thymocytes. Concanavalin A (Con A)-treated and untreated cells were extracted with 0.5% Triton X-100 and the remaining insoluble material was extracted under conditions allowing actin depolymerization. In the absence of Con A treatment, little T200 could be recovered from the depolymerized insoluble fraction. However, in T cells treated with capping concentrations of Con A, a considerable amount of T200 was rendered insoluble in nonionic detergent, and T200 could be recovered from the insoluble fraction by a buffer which dissociates actin polymers. A lesser, but still significant, amount of T200 associated with the detergent-insoluble fraction of thymocytes treated with concentrations of Con A and succinyl Con A, which are mitogenic for T cells. We also found that in T cells treated with mitogenic concentrations of succinyl Con A, more T200 associated with cytoskeleton than did H-2 or LFA-1 molecules. Because T200 is such a predominant molecule on the surface of T cells, such translocations of the molecule may have a major impact on the physiology of the cell, especially if T200 functions as a protein tyrosine phosphatase as recent evidence by others suggests.