Purification of opsonically active human and rat cold-insoluble globulin (plasma fibronectin)

Molnár, J.; Gelder, Frank B.; Zong, Lai Ming; Siefring, Gerald E.; Credo, R.B.; Lóránd, L.

doi:10.1021/bi00585a010

Cited by 67 publications

(55 citation statements)

References 34 publications

(51 reference statements)

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“…In agreement with others (16,33,40), we observed that fibronectin and heparin were required for the binding of gelatin-latex to liver slices (data not shown) . Whereas the data are consistent with the phagocytosis of these particles by Kupffer cells as suggested by others (3), this assay is unable to discriminate between ingestion and membrane attachment of these particles or between binding to hepatocytes and binding to other cells .…”

Section: Fibronectin Promotes the Binding Of The Latex Particles By Psupporting

confidence: 93%

“…Gelatin was conjugated to latex by a modification of the procedure of Molnar et al (33) . One part gelatin (10 mg/ml) in distilled water, two parts latex particles (Dow Diagnostics, Indianapolis, Ind.…”

Section: Preparation Of Gelatin-conjugated Latexmentioning

confidence: 99%

“…Initially, to compare our data with those of others, we used an assay in which slices of rat liver were incubated with the beads plus various serum preparations and heparin (33) . In agreement with others (16,33,40), we observed that fibronectin and heparin were required for the binding of gelatin-latex to liver slices (data not shown) .…”

Section: Fibronectin Promotes the Binding Of The Latex Particles By Pmentioning

confidence: 99%

“…Previous reports (33,40) had indicated a dependence on heparin of the liver slice assay, so we investigated its role in our assay system. If heparin was omitted from the incubation mixture, the level of binding in the presence of pFN approximated that of samples without pFN (Fig.…”

Section: Fibronectin Promotes the Binding Of The Latex Particles By Pmentioning

confidence: 99%

“…It also promotes the binding of particles to liver slices (12,16,31,40) and the uptake of gelatin-coated particles by rat peritoneal macrophages (13,18) . This a-2-globulin has recently been purified from rat and human sera and found to be immunologically and functionally identical with plasma fibronectin (2,33), a dimeric glycoprotein with subunits of 230,000 daltons (3) . Fibronectin is known to be involved in adhesion of cells to solid substrata and has specific binding sites for gelatin, glycosaminoglycans (i .e ., heparin), fibrinogen, and fibrin, and can be crosslinked to fibrin or collagen by Factor XIII (reviewed in references 17, 21, 35, 32 47, 49) .…”

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confidence: 99%

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Phagocytosis of gelatin-latex particles by a murine macrophage line is dependent on fibronectin and heparin.

Water¹,

Schroeder²,

Crenshaw³

et al. 1981

The Journal of Cell Biology

116

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It has been suggested that fibronectin plays a role in clearing particles from the circulation by promoting binding to phagocytes of the reticuloendothelial system . By use of a well-defined system to investigate the possible opsonic role of fibronectin, we have studied the uptake of gelatin-coated latex particles by a murine macrophage cell line (P388D 1 ) .Fibronectin promotes binding of gelatin-coated beads to these cells in both suspension and monolayer cultures . In both cases there is a requirement for heparin as a cofactor . Other glycosaminoglycans (chondroitin sulfates A and C, dermatan sulfate, and keratan sulfate) were inactive, whereas heparan sulfate was somewhat active .Proof that beads were actually endocytosed was obtained by electron microscopy, which showed beads internalized in membrane-bounded vesicles, and by immunofluorescence analyses, using antibodies to fibronectin to stain external beads. Two rapid assays for the opsonic activity of fibronectin were developed based on differential centrifugation of cellassociated beads and on the immunofluorescence procedure . Binding and endocytosis were time-and temperature-dependent and varied with the amount of gelatin on the beads and with the concentrations of fibronectin and heparin added, and could be inhibited by F(ab') 2 antifibronectin . These studies provide a sound basis for a detailed analysis of the interaction of fibronectin with the cell surface and of its involvement in endocytosis.Phagocytosis is the process by which particles (bacteria, damaged cells, fibrin aggregates, etc .) are bound and endocytosed by phagocytic cells . Recognition of these particles by the cells is due to the presence of humoral factors (opsonins) such as immunoglobulins or complement, which coat the particles and promote their phagocytosis (44). Recently, an a-2-globulin from plasma has been shown to facilitate clearance of certain particles from the circulation (31,32,42) . It also promotes the binding of particles to liver slices (12,16,31,40) and the uptake of gelatin-coated particles by rat peritoneal macrophages (13,18) . This a-2-globulin has recently been purified from rat and human sera and found to be immunologically and functionally identical with plasma fibronectin (2, 33), a dimeric glycoprotein with subunits of 230,000 daltons (3) . Fibronectin is known to be involved in adhesion of cells to solid substrata and has specific binding sites for gelatin, glycosaminoglycans (i .e ., heparin), fibrinogen, and fibrin, and can be crosslinked to fibrin or collagen by Factor XIII (reviewed in references 17, 21, 35, 32 47, 49) . Fibronectin has also been shown to bind to bacteria' (27, 36), but has not been shown to promote phagocytosis of bacteria .To define the conditions necessary for endocytosis of fibronectin-coated particles, we have studied an established mouse macrophagelike cell line (P388D,) by using a centrifugation assay, as well as electron and immunofluorescence microscopy. We find that endocytosis of gelatin-conjugated latex beads...

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Section: Fibronectin Promotes the Binding Of The Latex Particles By Psupporting

confidence: 93%

Section: Preparation Of Gelatin-conjugated Latexmentioning

confidence: 99%

Section: Fibronectin Promotes the Binding Of The Latex Particles By Pmentioning

confidence: 99%

Section: Fibronectin Promotes the Binding Of The Latex Particles By Pmentioning

confidence: 99%

mentioning

confidence: 99%

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Phagocytosis of gelatin-latex particles by a murine macrophage line is dependent on fibronectin and heparin.

Water¹,

Schroeder²,

Crenshaw³

et al. 1981

The Journal of Cell Biology

116

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Sequential Prospective Analysis of the Nonspecific Host Defense System After Thermal Injury

Deitch¹,

Gelder²,

McDonald³

1984

Arch Surg

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Affinity Peptidomics Approach to Protein Detection, Quantification, and Protein Affinity Assays: Application to Forensics and Biometrics

Bailes

Soloviev

2007

Peptidomics

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Purification of opsonically active human and rat cold-insoluble globulin (plasma fibronectin)

Cited by 67 publications

References 34 publications

Phagocytosis of gelatin-latex particles by a murine macrophage line is dependent on fibronectin and heparin.

Phagocytosis of gelatin-latex particles by a murine macrophage line is dependent on fibronectin and heparin.

Sequential Prospective Analysis of the Nonspecific Host Defense System After Thermal Injury

Affinity Peptidomics Approach to Protein Detection, Quantification, and Protein Affinity Assays: Application to Forensics and Biometrics

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