Multimeric analysis of von Willebrand factor in megakaryocytes

von Willebrand factor (vWF) is a glycoprotein that appears to play a major role in subserving the adhesion of platelets to subendothelium during hemostasis. Endothelial cells have been shown capable of synthesizing and releasing this large, multimeric glycoprotein that normally circulates in the plasma in association with the factor VIII coagulant molecule. Megakaryocytes, the precursor cells of blood platelets, also appear to possess vWF biosynthetic capacity, since cultured guinea pig megakaryocytes have been shown to produce a polypeptide precipitable by antibody to vWF. We now report a study of the multimeric structure of vWF in the megakaryocyte, as well as a quantitative comparison of megakaryocyte vWF with that of platelets and plasma in the guinea pig. Multimeric analysis on SDS agarose gels employing ¹²⁵I-emu anti-human vWF revealed striking homology between human and guinea pig vWF. Platelets and megakaryocytes from the same guinea pigs contained vWF of highly comparable multimeric composition. Moreover, megakaryocytes and platelets both contained a subset of very high molecular weight multimers not present in plasma. Quantitation of vWF in megakaryocytes and platelets was achieved with a radioimmunoassay performed on detergent (NP-40) lysates of washed cells. These measurements showed that megakaryocytes and platelets contain 0.079 and 0.069 U of vWF per mg protein, respectively. The results of these studies suggest that megakaryocytes represent the primary, if in fact not sole, source of platelet vWF.