title: Triplet structure of von Willebrand factor reflects proteolytic degradation of high molecular weight multimers.
abstract: High molecular weight (HMW) and low molecular weight (LMW) forms of von Willebrand factor (vWF) were isolated from normal human plasma in the presence of protease inhibitors. HMW and LMW vWF preparations were subjected to reduction of interdimeric disulfide bridges under mild reducing conditions. Following sodium dodecyl sulfate electrophoresis in 3% agarose, the vWF bands were detected by immunoblotting with a polyclonal rabbit anti-vWF antiserum as well as with two monoclonal antibodies directed against epitopes located in the NH2-terminal (MAb 418) or in the COOH-terminal (MAb 9) region of the vWF subunit. Our results suggest that the slowest migrating band of the dimeric triplet set of LMW vWF represents an asymmetric structure composed of an intact subunit to which one NH2-terminal and one COOH-terminal fragment are linked by disulfide bridges. The intermediate band of the first triplet of LMW vWF strongly reacted with MAb 9 but not with MAb 418, indicating that it represents a dimer of COOH-terminal fragments. The fastest migrating band of the same triplet is apparently a dimer of the NH2-terminal fragments because it reacted with MAb 418 but not with MAb 9. Each next higher family of triplets seems to contain one more asymmetric fragment of dimeric size. These results are compatible with a model according to which LMW forms of vWF are derived from HMW vWF by proteolytic cleavage in the circulating blood.
authors.affiliation: Central Hematology Laboratory, Inselspital, University of Bern, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Furlan
authors.forename: R
authors.initials: R
authors.lastname: Robles
authors.forename: D
authors.initials: D
authors.lastname: Affolter
authors.forename: D
authors.initials: D
authors.lastname: Meyer
authors.forename: P
authors.initials: P
authors.lastname: Baillod
authors.forename: B
authors.initials: B
authors.lastname: Lämmle
mesh_terms: D002850:Chromatography, Gel; D004587:Electrophoresis, Agar Gel; D006801:Humans; D015151:Immunoblotting; D046911:Macromolecular Substances; D008970:Molecular Weight; D014841:von Willebrand Factor / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D046911:Macromolecular Substances; D014841:von Willebrand Factor