+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Expression of variant von willebrand factor vwf complementary dna in heterologous cells requirement of the pro polypeptide in vwf multimer formation

EMBO (European Molecular Biology Organization) Journal 6(10): 2885-2890

Expression of variant von willebrand factor vwf complementary dna in heterologous cells requirement of the pro polypeptide in vwf multimer formation

Von Willebrand factor (vWF) is a multimeric plasma glycoprotein synthesized by vascular endothelial cells as a pre-pro-polypeptide with a highly repetitive domain structure, symbolized by the formula: (H)-D1-D2-D'-D3-A1-A2-A3-D4-B1-B2-B3-C1-C2-(OH) a heterologous expression system for the synthesis of recombinant vWF protein was developed, consisting of a monkey kidney cell line (COS-1), transfected with full-length vWF cDNA. This system was shown to mimic the constitutive secretory pathway of vWF in endothelial cells, since dimerization and multimerization occur similarly. To determine whether the pro-polypeptide, composed of the domains D1 and D2, is involved in vWF multimerization, a vWF cDNA was constructed that lacked the coding sequence for the pro-polypeptide. The mutant vWF protein, expressed by COS-1 cells transfected with this cDNA, did not assemble beyond the dimer stage. From this observation, we conclude that (i) dimerization does not involve the pro-polypeptide of pro-vWF and (ii) the presence of the pro-polypeptide, as part of pro-vWF, is obligatory for multimerization. It is argued that the interactions, required for interchain binding, are mediated by the D domains.

Accession: 005432398

Related references

Expression of variant von Willebrand factor (vWF) cDNA in heterologous cells: requirement of the pro-polypeptide in vWF multimer formation. Embo Journal 6(10): 2885-2890, 1987

Von willebrand factor vwf pro polypeptide is required for vwf multimer formation. Thrombosis & Haemostasis 58(1): 8, 1987

The pro-polypeptide of von Willebrand factor is required for the formation of a functional factor VIII-binding site on mature von Willebrand factor. Biochemical Journal 274: 257-261, 1991

Proteolytic cleavage of the precursor of von Willebrand factor is not essential for multimer formation. Journal of Biological Chemistry 263(17): 7921-7924, 1988

Multicenter comparison of von Willebrand factor multimer sizing techniques. Report of the Factor VIII and von Willebrand Factor Subcommittee. Thrombosis and Haemostasis 54(4): 873-876, 1985

Structure of pre pro von willebrand factor and its expression in heterologous cells. Nature (London) 324(6094): 270-273, 1986

Characterization of factor VIII/von Willebrand factor concentrates using a modified method of von Willebrand factor multimer analysis. Haemophilia 4 Suppl 3: 25-32, 1999

Evaluation of monoclonal antibodies to von willebrand factor antigen for use in autoradiography von willebrand factor multimer analyses. Ricerca in Clinica e in Laboratorio 16(1): 95, 1986

Novel mutations in types 2 & 3 von Willebrand disease and correlation with von Willebrand factor multimer patterns. 2007

Lack of multimer organization of von Willebrand factor in an acquired von Willebrand syndrome. British Journal of Haematology 116(4): 899-904, March, 2002