Mobility and distribution of a cell surface glycoprotein and its interaction with other membrane components

Schlessinger, Joseph; Barak, LS; Hammes, Gordon G.; Yamada, Katsushi; Pastan, Ira; Webb, Watt W.; Elson, Elliot L.

doi:10.1073/pnas.74.7.2909

Cited by 83 publications

(27 citation statements)

References 17 publications

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“…FN crosslinks with itself and other proteins through disulfide links (9,45). FN on cell surfaces is known to be immobile and may serve as an anchorage point for cell attachment (44). Thus, FN in the subendothelium may be an important structural component which plays a role in endothelial cell attachment and adhesiveness under normal conditions and after injury to the vascular wall.…”

Section: Discussionmentioning

confidence: 99%

“…Fibroblast cultures contain fibrils which stain for both FN and collagen when examined by immunofluorescence microscopy (17). The basis for this congruence of location may be the recently described noncovalent interactions of FN with other FN molecules (44) and with collagen (18). FN crosslinks with itself and other proteins through disulfide links (9,45).…”

Section: Discussionmentioning

confidence: 99%

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Synthesis of fibronectin by cultured human endothelial cells.

Jaffe¹,

Mosher²

1978

The Journal of Experimental Medicine

448

113

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Plasma fibronectin is probably the major nonimmune particulate opsonin in blood and is cross-linked to fibrin during the final stage of blood coagulation. Fibronectin also occurs in an insoluble form in basement membranes especially those underlying endothelial cells and in loose connective tissue. Fibronectin was demonstrated in cultured human endothelial cells and in the surrounding extracellular matrix by immunofluorescence microscopy by using antibody to human plasma fibronectin. Cultured human endothelial cells released fibronectin into the culture medium which was immunologically identical to the fibronectin in human plasma. Cultured human endothelial cells were labeled with [3H] leucine. The radioactive fibronectin present in the endothelial postculture medium and in urea extracts of cellular monolayers was isolated with either anti-fibronectin coupled to Protein A-Sepharose or double antibody immunoprecipitation and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When reduced, the [3H] fibronectin synthesized by cultured endothelial cells had the same mol wt (approximately 200,000) as plasma fibronectin. Unreduced, the [3H] fibronectin synthesized by endothelial cells migrated as a dimer, as did plasma fibronectin. Fibronectin accounted for approximately 15% of the protein synthesized and released by endothelial cells into the culture medium. Thus, cultured endothelial cells synthesize fibronectin, secrete it into the culture medium, and incorporate it into extracellular matrix. The results suggest that the endothelial cell is potentially a major site of synthesis of circulating plasma fibronectin. In addition, fibronectin derived from endothelial cells may be an important structural component of the subendothelium.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Synthesis of fibronectin by cultured human endothelial cells.

Jaffe¹,

Mosher²

1978

The Journal of Experimental Medicine

448

113

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“…It has been shown that fibronectin fibers do not impede the diffusion of lipid probes and cell surface antigens [28]. Concanavalin A, however, is immobilized by binding directly to immobile fibronectin fibers m1.…”

Section: Modulation Of Lipid and Protein Mobilitiesmentioning

confidence: 99%

Analysis of cell surface interactions by measurements of lateral mobility

Elson

Reidler

1979

J Supramol Struct

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Interactions of cell surface components with one another and with structures inside and outside the cell may have important physiological functions in the transmission of signals and the assembly of specialized structures. These interactions may be detected and analyzed through their effects on the lateral mobility of cell surface molecules. Measurements by a fluorescence photobleaching method have shown that in general lipid-like molecules diffuse rapidly and freely through the plasma membrane, whereas proteins move much more slowly or appear to be immobile. This dichotomy has been supposed t o result from forces beyond the viscosity of the lipid bilayer, which specifically retard the diffusion of membrane proteins. This general picture should be qualified, however, by noting that the lateral mobility of lipid-like molecules can be influenced in detail by changes in the state of the plasma membrane such as result from mitosis or fertilization. The interactions of cell surface proteins that limit their lateral mobility are unknown. The effects of binding concanavalin A t o localized regions of cell surface show that these interactions can vary in subtle and complex ways. It may soon be useful t o interpret mobility experiments in terms of simple reaction models that attempt to describe surface interactions in physicochemical terms. More experimental data are needed to carry out this program and to relate interactions that affect mobility to the structural connections between cell surface components and the cytoskeleton, which have been detected by biochemical methods and electron and irnmunofluorescence microscopy.

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“…For example, the acetylcholine receptor (AChR) localized at neuromuscular junctions and in patches on myotubes (7,8), and frbronectin on the surface of cultured fibroblasts (9) .…”

mentioning

confidence: 99%

Enhanced molecular diffusibility in muscle membrane blebs: release of lateral constraints.

Tank¹,

Wu²,

Webb³

1982

The Journal of Cell Biology

249

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Measurements of lateral molecular diffusion on blebs formed on the surfaces of isolated muscle cells and myoblasts are reported . These blebbed membranes retain integral proteins but apparently separate from the detectable cytoskeleton . On blebs, acetylcholine receptors, concanavalin A receptors, and stearoyldextran extrinsic model receptor molecules are free to diffuse with a diffusion coefficient (D) -3 x 10-9 cm'/s, which is close to the value predicted for hydrodynamic drag in the lipid membrane . In contrast, diffusion of these typical receptors in intact cell membranes is constrained to D :Z 10-10 cm2/s with substantial fractions virtually nondiffusible (D < 10-' 2 cm2/s) . Lipid analog diffusion is also slightly enhanced in blebs as expected of evanescent lipid protein interaction . This strong enhancement of membrane protein diffusion is attributed to release from unidentified natural constraints that is induced in some way by detachment of the bleb membrane .The lateral diffusion of plasma membrane components has become routinely accessible to measurement by fluorescence photobleaching recovery (FPR) (1) . Taken together, data accumulated for vertebrate cells in culture establish several distinctive common characteristics of lateral diffusibility of cell surface components (2--6) : (a) A substantial fraction of the population of each cell surface protein is not detectably diffusible over distances of several micrometers during an experimental interval ofhours . Thus the diffusible fraction (R) is said to be <100%, and it is often <75%. (b) Diffusion coefficients (D) of the diffusible fraction ofcell membrane proteins do not generally exceed 1 x 10-1°cm 2/s and are frequently much smaller. (Implied diffusive transit time across a 10-8m cell: 1/2 h.) (c) Plasma membrane lipid diffusibility is virtually complete over distances of several micrometers (R = 100%, except possibly in a few compartmentalized cell types) with diffusion coefficients of the order of 1 x 10-e cm2/s, i.e., usually >100 times faster than membrane proteins. (Implied diffusive transit time across a 10-N.m cell: 1/2 min.)Certain proteins are virtually nondiffusible in toto (R = 0%) on the cell surface. For example, the acetylcholine receptor (AChR) localized at neuromuscular junctions and in patches on myotubes (7,8), and frbronectin on the surface of cultured fibroblasts (9) .In contrast with the cell surface, protein molecules that are reconstituted into phospholipid model membranes diffuse nearly as fast as lipid molecules and with no immobile fraction.

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Mobility and distribution of a cell surface glycoprotein and its interaction with other membrane components

Cited by 83 publications

References 17 publications

Synthesis of fibronectin by cultured human endothelial cells.

Synthesis of fibronectin by cultured human endothelial cells.

Analysis of cell surface interactions by measurements of lateral mobility

Enhanced molecular diffusibility in muscle membrane blebs: release of lateral constraints.

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