Measurement of the lateral mobility of cell surface components in single, living cells by fluorescence recovery after photobleaching.

The use of fluorescence recovery after photobleaching (FRAP) techniques to monitor the lateral mobility of plant lectin-receptor complexes on the surface of single, living mammalian cells is described in detail. FRAP measurements indicate that over 75% of the wheat germ agglutinin receptor (WGA-receptor) complexes on the surface of human embryo fibroblasts are mobile. These WGS-receptor complexes diffuse laterally (as opposed to flow) on the cell surface with a diffusion coefficient in the range of 2 X 10(-11) to 2 X 10(-10) cm2/sec. Both the percentage of mobile WGA-receptor complexes and the mean diffusion coefficient of these complexes are higher than that obtained from earlier FRAP measurements of the mobility of concanavalin A-receptor (Con A-receptor) complexes in a variety of cell types. The possible reasons for the differing mobilities of WGA and Con A receptors are discussed.

[1]  W. Webb,et al.  Lateral transport on cell membranes: mobility of concanavalin A receptors on myoblasts. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[2]  W. Webb,et al.  Lateral transport of a lipid probe and labeled proteins on a cell membrane. , 1977, Science.

[3]  G. Nicolson,et al.  The interactions of lectins with animal cell surfaces. , 1974, International review of cytology.

[4]  S. D. Petris,et al.  Concanavalin A receptors, immunoglobulins, and theta antigen of the lymphocyte surface. Interactions with concanavalin A and with Cytoplasmic structures. , 1975 .

[5]  M Edidin,et al.  Rotational and translational diffusion in membranes. , 1974, Annual review of biophysics and bioengineering.

[6]  W. Webb,et al.  Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. , 1976, Biophysical journal.

[7]  G. Nicolson,et al.  Local anesthetics affect transmembrane cytoskeletal control of mobility and distribution of cell surface receptors. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[8]  The determination of local cell membrane diffusion coefficients. , 1976, Journal of theoretical biology.

[9]  K. Jacobson,et al.  Measurement of the translational mobility of concanavalin A in glycerol-saline solutions and on the cell surface by fluorescence recovery after photobleaching. , 1976, Biochimica et biophysica acta.

[10]  S. H. Thompson,et al.  Fluorescent antibody staining. 3. Preparation of fluorescein-isothiocyanate-labeled antibodies. , 1965, Journal of immunology.

[11]  G M Edelman,et al.  Surface modulation in cell recognition and cell growth. , 1976, Science.

[12]  L D Dickson,et al.  Characteristics of a propagating gaussian beam. , 1970, Applied optics.

[13]  M. Karnovsky,et al.  EFFECTS OF COLCHICINE, CYTOCHALASIN B, AND 2-DEOXYGLUCOSE ON THE TOPOGRAPHICAL ORGANIZATION OF SURFACE-BOUND CONCANAVALIN A IN NORMAL AND TRANSFORMED FIBROBLASTS , 1974, The Journal of cell biology.

[14]  M. Bretscher,et al.  Mammalian plasma membranes , 1975, Nature.

[15]  M. Edidin,et al.  Lateral diffusion of concanavalin A receptors in the plasma membrane of mouse fibroblasts. , 1976, Biochimica et biophysica acta.

[16]  G. Nicolson,et al.  The cancer cell: dynamic aspects and modifications in cell-surface organization (first of two parts). , 1976, The New England journal of medicine.

[17]  Mu-ming Poo,et al.  Lateral diffusion of rhodopsin in the photoreceptor membrane , 1974, Nature.

[18]  G. Entine,et al.  Lateral Diffusion of Visual Pigment in Photoreceptor Disk Membranes , 1974, Science.

[19]  Y. Suzaki,et al.  Measurement of the microm sized radius of Gaussian laser beam using the scanning knife-edge. , 1975, Applied optics.

[20]  R. Peters,et al.  A microfluorimetric study of translational diffusion in erythrocyte membranes. , 1974, Biochimica et biophysica acta.

[21]  P. Greenaway,et al.  The purification and characterization of wheat-germ agglutinin. , 1972, The Biochemical journal.

[22]  K. Jacobson,et al.  Effects of local anesthetics on membrane properties. II. Enhancement of the susceptibility of mammalian cells to agglutination by plant lectins. , 1975, Biochimica et biophysica acta.

[23]  M Edidin,et al.  Measurement of membrane protein lateral diffusion in single cells. , 2003, Science.