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. 1979 Nov;64(5):1298–1302. doi: 10.1172/JCI109585

Carbohydrate of the Factor VIII/von Willebrand Factor in von Willebrand's Disease

T S Zimmerman 1, R Voss 1, T S Edgington 1
PMCID: PMC371276  PMID: 315413

Abstract

We have examined the plasma Factor VIII/von Willebrand factor (FVIII/vWF) molecule from 16 patients with von Willebrand's disease, and have found no evidence of a significant decrease of carbohydrate content in 15 of these patients. FVIII/vWF was isolated by preparative counter immunoelectrophoresis directly from plasma using antibody to Factor VIII-related antigen, reduced in sodium dodecyl sulfate in the presence of urea, and electrophoresed in 5% polyacrylamide gels to separate the FVIII/vWF subunit from other proteins. Duplicate gels were stained by either the periodic acid-Schiff (PAS) reaction or by Coomassie Brilliant Blue G250. The ratio of Coomassie: PAS was determined by spectrophotometric scanning of the gels. Transferrin was used as an internal reference standard. The ratio for 23 normal individuals was 2.4±0.38 and the observed range was 1.8-3.8. 15 patients with von Willebrand's disease fell within this range. One patient independently reported as having decreased FVIII/vWF carbohydrate was also studied by this technique. A ratio of 6.8 was found, indicative of decreased, though not absent, carbohydrate. Cold insoluble globulin did not represent a significant contaminant in these analyses. 11 of the von Willebrand's disease patients with normal FVIII/vWF carbohydrate had abnormal crossed immunoelectrophoretic patterns characterized by absence of the less anodic forms of Factor VIII-related antigen. Four patients had normal patterns. These studies indicate that an absence or decrease of PAS reactive FVIII/vWF carbohydrate is not a consistent abnormality in von Willebrand's disease.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Engvall E., Ruoslahti E. Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int J Cancer. 1977 Jul 15;20(1):1–5. doi: 10.1002/ijc.2910200102. [DOI] [PubMed] [Google Scholar]
  2. Gralnick H. R., Coller B. S., Sultan Y. Carbohydrate deficiency of the factor VIII/von Willebrand factor Protein in von Willebrand's disease variants. Science. 1976 Apr 2;192(4234):56–59. doi: 10.1126/science.1083071. [DOI] [PubMed] [Google Scholar]
  3. Gralnick H. R. Factor VIII/von Willebrand factor protein. Galactose a cryptic determinant of von Willebrand factor activity. J Clin Invest. 1978 Aug;62(2):496–499. doi: 10.1172/JCI109152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gralnick H. R., Sultan Y., Coller B. S. Von Willebrand's disease: combined qualitative and quantitative abnormalities. N Engl J Med. 1977 May 5;296(18):1024–1030. doi: 10.1056/NEJM197705052961802. [DOI] [PubMed] [Google Scholar]
  5. Kapitany R. A., Zebrowski E. J. A high resolution PAS stain for polyacrylamide gel electrophoresis. Anal Biochem. 1973 Dec;56(2):361–369. doi: 10.1016/0003-2697(73)90202-9. [DOI] [PubMed] [Google Scholar]
  6. Koutts J., Walsh P. N., Plow E. F., Fenton J. W., 2nd, Bouma B. N., Zimmerman T. S. Active release of human platelet factor VIII-related antigen by adenosine diphosphate, collagen, and thrombin. J Clin Invest. 1978 Dec;62(6):1255–1263. doi: 10.1172/JCI109246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  9. Montgomery R. R., Zimmerman T. S. von Willebrand's disease antigen II. A new plasma and platelet antigen deficient in severe von Willebrand's disease. J Clin Invest. 1978 Jun;61(6):1498–1507. doi: 10.1172/JCI109070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Over J., Bouma B. N., van Mourik J. A., Sixma J. J., Vlooswijk R., Bakker-Woudenberg I. Heterogeneity of human factor VIII. I. Characterization of factor VIII present in the supernatant of cryoprecipitate. J Lab Clin Med. 1978 Jan;91(1):32–46. [PubMed] [Google Scholar]
  11. Reisner A. H., Nemes P., Bucholtz C. The use of Coomassie Brilliant Blue G250 perchloric acid solution for staining in electrophoresis and isoelectric focusing on polyacrylamide gels. Anal Biochem. 1975 Apr;64(2):509–516. doi: 10.1016/0003-2697(75)90461-3. [DOI] [PubMed] [Google Scholar]
  12. Sodetz J. M., Paulson J. C., Pizzo S. V., McKee P. A. Carbohydrate on human factor VIII/von Willebrand factor. Impairment of function by removal of specific galactose residues. J Biol Chem. 1978 Oct 25;253(20):7202–7206. [PubMed] [Google Scholar]
  13. Sodetz J. M., Pizzo S. V., McKee P. A. Relationship of sialic acid to function and in vivo survival of human factor VIII/von Willebrand factor protein. J Biol Chem. 1977 Aug 10;252(15):5538–5546. [PubMed] [Google Scholar]
  14. Zimmerman T. S., Hoyer L. W., Dickson L., Edgington T. S. Determination of the von Willebrand's disease antigen (factor VIII-related antigen) in plasma by quantitative immunoelectrophoresis. J Lab Clin Med. 1975 Jul;86(1):152–159. [PubMed] [Google Scholar]

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