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Measurement of the translation mobility of concanavalin a in glycerol-saline solutions and on the cell surface by fluorescence recovery after photobleaching

1976; Elsevier BV; Volume: 433; Issue: 1 Linguagem: Inglês

10.1016/0005-2736(76)90189-9

1879-2642

Kenneth A. Jacobson, E. S. Wu, George Poste,

Protein Interaction Studies and Fluorescence Analysis

The fluorescence recovery kinetics of succinyl-fluorescein Concanavalin A (S-F-ConA) in glycerol-physiological saline solutions of high viscosity and when bound to the surface of mouse fibroblast were measured following brief photobleaching using a laser excited fluorescence microscope. In the high viscosity solutions, the recovery kinetics, interpreted on the basis of a simple diffusion model, yielded a diffusion coefficient in close agreement with the values predicted by the Stokes-Einstein equation. Recovery kinetics for S-F-ConA bound to the surface of mouse 3T3 and SV3T3 cells cultured in vitro yielded diffusion coefficients in the range of 5–10·10−11cm2/s, values considerably lower than those reported previously for membrane protiens. These measurement indicated that a considerable fraction of the S-F-ConA molecules bound to the cell surface are immobilized. These results are discussed in relation to current concepts of lateral motion of protein components within natural membranes.