Requirement for Both D Domains of the Propolypeptide in von Willebrand Factor Muitimerization and Storage

Summary

Biosynthesis of the adhesive glycoprotein von Willebrand factor (vWf) by endothelial cells results in constitutive secretion of small multimers and storage of the largest multimers in rodshaped granules called Weibel-Palade bodies. This pattern is reproduced by expression of pro-vWf in heterologous cells with a regulated pathway of secretion, that store the recombinant protein in similar elongated granules. In these cells, deletion of the vWf prosequence prevents vWf storage. The prosequence, composed of two homologous domains (D1 and D2), actively participates in vWf multimer formation as well. We expressed deletion mutants lacking either the D1 domain (D2vWf) or the D2 domain (D1vWf) in various cell lines to analyze the relative importance of each domain in vWf muitimerization and storage. Both proteins were secreted efficiently without being retained in the endoplasmic reticulum. Despite this, neither multimerized past the dimer stage and they were not stored. We conclude that several segments of the prosequence are jointly involved in vWf muitimerization and storage.

• References

• 1 Wagner DD. Cell biology of von Willebrand factor. Annu Rev Cell Biol 1990; 6: 217-246
• 2 Ruggeri ZM, Zimmerman TS. von Willebrand factor and von Willebrand disease. Blood 1987; 70: 895-904
• 3 Nachman R, Levine R, Jaffe EA. Synthesis of factor VIII antigen by cultured guinea pig megakaryocytes. J Clin Invest 1977; 60: 914-921
• 4 Jaffe EA, Hoyer LW, Nachman RL. Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J Clin Invest 1973; 52: 2757-2764
• 5 Mayadas TN, Wagner DD. von Willebrand factor biosynthesis and processing. Ann of NY Acad of Sci 1991; 614: 153-166
• 6 Weibel ER, Palade GE. New cytoplasmic components in arterial endothelia. J Cell Biol 1964; 23: 101-112
• 7 Wagner DD, Olmsted JB, Marder VJ. Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol 1982; 95: 355-360
• 8 Bonfanti R, Furie BC, Furie B, Wagner DD. PADGEM (GMP140) is a component of Weibel-Palade bodies of human endothelial cells. Blood 1989; 73: 1109-1112
• 9 McEver RP, Beckstead JH, Moore KL, Marshall-Carlson L, Bainton DF. GMP-140, a platelet alpha-granule membrane protein, is also synthesized by vascular endothelial cells and is localized in Weibel-Palade bodies. J Clin Invest 1989; 84: 92-99
• 10 Levine JD, Harlan JM, Harker LA, Joseph ML, Counts RB. Thrombin-mediated release of factor VIII antigen from human umbilical vein endothelial cells in culture. Blood 1982; 60: 531-534
• 11 Sporn LA, Marder VJ, Wagner DD. Inducible secretion of large biologically potent von Willebrand factor multimers. Cell 1986; 46: 185-190
• 12 Ewenstein BM, Warhol MJ, Handin RI, Pober JS. Composition of the von Willebrand factor storage organelle (Weibel-Palade body) isolated from cultured human umbilical vein endothelial cells. J Cell Biol 1987; 104: 1423-1433
• 13 Sadler JE. The molecular biology of human von Willebrand factor. In: Coagulation and Bleeding Disorders: The Role of Factor VIII and von Willebrand factor. Zimmerman TS, Ruggeri ZM. ed. New York: Marcel Dekker Inc; 1989. 9. 117-136
• 14 Verweij CL, Hart M, Pannekoek H. Expression of variant von Willebrand factor (vWf) cDNA in heterologous cells: Requirement of the propolypeptide in vWf multimer formation. EMBO J 1987; 6: 2885-2890
• 15 Wise RJ, Pittman DD, Handin RI, Kaufman RJ, Orkin SH. The propeptide of von Willebrand factor independently mediates the assembly of von Willebrand multimers. Cell 1988; 52: 229-236
• 16 Wagner DD, Saffaripour S, Bonfanti R, Sadler JE, Cramer EM, Chapman B, Mayadas TN. Induction of specific storage organelles by von Willebrand factor propolypeptide. Cell 1991; 64: 403-413
• 17 Voorberg J, Fontjin R, Calafat J, Janssen H, van Mourik JA, Pannekoek H. Biogenesis of von Willebrand factor-containing organelles in heterologous transfected CV-1 cells. EMBO J 1993; 12: 749-758
• 18 Voorberg J, Fontjin R, van Mourik JA, Pannekoek H. Domains involved in multimer assembly of von Willebrand factor (vWf): multimerization is independent of dimerization. EMBO J 1990; 9: 797-803
• 19 Bonthron DT, Handin RI, Kaufman RJ, Wasley LC, Orr EC, Mitsock LM, Ewenstein B, Loscalzo J, Ginsburg D, Orkin SH. Structure of pre-pro-von Willebrand factor and its expression in heterologous cells. Nature 1986; 324: 270-273
• 20 Journet AM, Saffaripour S, Martin CramerE, Tenza D, Wagner DD. von Willebrand factor storage requires intact prosequence cleavage site. Eur J Cell Biol 1993; 60: 31-41
• 21 Bonthron D, Orr EC, Mitsock LM, Ginsburg D, Handin RI, Orkin SH. Nucleotide sequence of pre-pro-von Willebrand factor cDNA. Nucl Acids Res 1986; 14: 7125-7127
• 22 Mayadas T, Wagner DD. In vitro multimerization of von Willebrand factor is triggered by low pH. J Biol Chem 1989; 264: 13497-13503
• 23 Wagner DD, Marder VJ. Biosynthesis of von Willebrand protein by human endothelial cells. Identification of a large precursor polypeptide chain. J Biol Chem 1983; 258: 2065-2067
• 24 Wagner DD, Lawrence SO, Ohlsson-Wilhelm BM, Fay PJ, Marder VJ. Topology and order of formation of interchain disulfide bonds in von Willebrand factor. Blood 1987; 69: 27-32
• 25 Wagner DD, Mayadas T, Marder VJ. Initial glycosylation and acidic pH in the Golgi apparatus are required for multimerization of von Willebrand factor. J Cell Biol 1986; 102: 1320-1324
• 26 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
• 27 Verweij CL, Hart M, Pannekoek H. Proteolytic cleavage of the precursor of von Willebrand factor is not essential for multimer formation. J Biol Chem 1988; 263: 7921-7924
• 28 Wagner DD, Marder VJ. Biosynthesis of von Willebrand protein by human endothelial cells: Processing steps and their intracellular localization. J Cell Biol 1984; 99: 2123-2130
• 29 Bentley A, Rees DJG, Rizza C, Brownlee GG. Defective propeptide processing of blood clotting factor IX caused by mutation of arginine to glutamine at position -4. Cell 1986; 45: 343-348
• 30 Rehemtulla A, Kaufman RJ. Preferred sequence requirements for cleavage of pro-von Willebrand factor by propeptide-processing enzymes. Blood 1992; 79: 2349-2355
• 31 Wagner DD, Fay PJ, Spom LA, Sinha S, Lawrence SO, Marder VJ. Divergent fates of von Willebrand factor and its propolypeptide (von Willebrand antigen II) after secretion from endothelial cells. Proc Natl Acad Sci USA 1987; 84: 1955-1959
• 32 Mayadas TN, Wagner DD. Vicinal cysteines in the prosequence play a role in von Willebrand factor multimer assembly. Proc Natl Acad Sci USA 1992; 89: 3531-3535