Optimization of a recombinant von Willebrand factor fragment as an antagonist of the platelet glycoprotein Ib receptor.

The binding of von Willebrand factor (vWF) to platelet glycoprotein (GP) Ib receptor is one of the initial events in thrombus formation. Previous studies have shown that RG12986, a reduced and alkylated recombinant fragment of vWF (Ser445-Val733), can inhibit binding of native vWF to GP Ib and offers potential as an anti-thrombotic agent. We have now evaluated a series of deletion mutants of RG12986 and found that reduced and alkylated rvWF508-704 is close to the minimal sequence with optimal RG12986-like activity (IC50 for inhibition of GP Ib-dependent platelet aggregation in the absence of modulators: 0.

Production and functional characterization of a recombinant fragment of von Willebrand factor (vWF): an antagonist to platelet receptor GP Ib.

We expressed a recombinant peptide fragment (Ser445-Val733) of human von Willebrand factor (vWF), containing the binding domain for the platelet receptor of GP Ib, in E. coli. This 33 kD peptide blocks binding of the intact vWF molecule to GP Ib in the presence of modulators.

Uptake of low density lipoprotein, albumin, and water by deendothelialized in vitro minipig aorta.

The purpose of this research was to study the effect of pressure on arterial hydration in vitro and the effect of pressure and flow (stirred reagent) on the in vitro transport of 125I-albumin and 125I-LDL into deendothelialized minipig aortas over a 24-hour period. It was found that the arterial hydration (fractional mass of water) was 0.740 +/- 0.

Effect of serum and stirring on diffusive 125I-albumin and Evans blue dye uptake.

The diffusive in vitro uptake of homologous 125I-albumin (MA, nmol.cm-2) and Evans blue dye (EBD) (ME, nmol.cm-2) by the deendothelialized canine aorta from serum and from a simple albumin solution with and without EBD and with and without vigorous stirring was measured in 18 preparations.

Cholesterol transfer from normal and atherogenic low density lipoproteins to Mycoplasma membranes.

The purpose of this study was to determine whether the free cholesterol of hypercholesterolemic low density lipoprotein from cholesterol-fed nonhuman primates has a greater potential for surface transfer to cell membranes than does the free cholesterol of normal low density lipoprotein. The low density lipoproteins were isolated from normal and hypercholesterolemic rhesus and cynomolgus monkeys, incubated with membranes from Acholeplasma laidlawii, a mycoplasma species devoid of cholesterol in its membranes, and the mass transfer of free cholesterol determined by measuring membrane cholesterol content. Since these membranes neither synthesize nor esterify cholesterol, nor degrade the protein or cholesterol ester moieties of low density lipoprotein, they are an ideal model with which to study differences in the cholesterol transfer potential of low density lipoprotein independent of the uptake of the intact low density lipoprotein particle.