The binding of purified factor VIII/Von Willebrand factor to collagens of differing type and form

Structural basis of von Willebrand factor binding to platelet glycoprotein Ib and collagen. Effects of disulfide reduction and limited proteolysis of polymeric von Willebrand factor.

“…Although vWF is multifunctional and binds to platelet GpIb (5,6,25,26), GpIIb-IIIa (30)(31)(32), components of the subendothelium such as collagen (7)(8)(9)16) and Factor VIII:C (28,33), the domain on native vWF polymer that binds to each of these receptors or proteins has not been localized. One our studies indicate (Fig.…”

“…Ristocetin was obtained from Bio/Data Corp., Hatboro, PA. Bovine serum albumin, polyethylene glycol M, 8,000 and M, 20,000, beta-mercaptoethanol, dextran sulfate M, 500,000, aprotinin, and phenylmethylsulfonyl fluoride (PMSF), were obtained from Sigma Chemical Co., St. Louis, MO. Staphylococcal V8 protease was obtained from Miles Laboratories, Inc., Naperville, IL.…”

“…5 liters of plasma obtained within 2-3 d of collection from the American Red Cross Blood Services-Northeast Region (Boston, MA), frozen at -20'C for 24 h, and thawed at 4VC for 18 h was used as the starting material. The resulting cryoprecipitate was collected by centrifugation at 6,500 g for 20 min, resuspended in 20 mM Na citrate, pH 6.1, and reprecipitated twice with final concentrations of 5 and 12% polyethylene glycol (M, 8,000) (PEG) as previously described.…”

“…Abbreviations used in this paper: GpIb, glycoprotein Ib vessel wall (1)(2)(3). Although the mechanism by which vWF promotes adhesion has not been fully delineated, there is evidence that vWF links platelets to the vessel wall by binding to platelet glycoprotein Ib (GpIb), as well as to exposed subendothelial collagen (4)(5)(6)(7)(8)(9). In previous studies, we, and others, have demonstrated vWF binding to GpIb and competition for binding by a soluble proteolytic fragment of GpIb, glycocalicin (10, 1 1).…”

Synthesis and expression of the fibroblast fibronectin receptor in human monocytes.

“…Sterile glass tissue-chamber slides (Lab-Tek 4802; Nunc, Inc., Naperville, IL) were pretreated with gelatin (2 mg/ml, Bio-Rad Laboratories, Richmond, CA) at room temperature for 2 h to block the nonspecific adherence ofthe monocytes to glass. After removal ofthe excess gelatin by aspiration, we added fibronectin, vitronectin, or von Willebrand factor, each of which binds to gelatin (19)(20)(21), and incubated the slides for 1 h at room temperature. After removal of the excess, unbound adhesive proteins, 1 ml of the labeled monocytes was added to each chamber ofthe tissue-chamber slides and incubated for 90 min at 370C in a 5% CO2 incubator.…”