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Electron microscopy of human factor VIII/von willebrand glycoprotein: Effect of reducing reagents on structure and function

Publication ,  Journal Article
Ohmori, K; Fretto, LJ; Harrison, RL; Switzer, MEP; Erickson, HP; McKee, PA
Published in: Journal of Cell Biology
November 1, 1982

The structure of native and progressively reduced human factor Vlll/von Willebrand factor (FVlll/vWF) was examined by electron microscopy and SDS gel electrophoresis and then correlated with its biological activities. Highly resolved electron micrographs of well-spaced, rotary-shadowed FVlII/vWF molecules showed their structure to consist of a very flexible filament that contains irregularly spaced small nodules. Filaments ranged from 50 to 1,150 nm with a mean length of 478 nm and lacked fixed, large globular domains as seen in fibrinogen and IgM. A population of multimeric FVlII/vWF species ranging in molecular weight from 1 to 5 million daltons and differing in size alternately by one and two subunits was observed on SDS-2% polyacrylamide-0.5% agarose gel electrophoresis. With progressive reduction of disulfide bonds by dithiothreitol (DTT), the electron microscopic size of FVIII/vWF decreased in parallel with increased electrophoretic mobility on SDS-agarose gels; between 0.1 and 0.5 mM DTT its structure changed from predominantly fibrillar species to large nodular forms. A 50% loss of vWF specific activity and FVIII procoagulant activity occurred at 0.4 mM DTT and 1 mM DTT, respectively, corresponding to the reduction of 4 and 12 disulfide bonds of the 62 disulfides per 200,000-dalton subunit. We conclude that reduction of a few critical disulfide bonds results in a major structural change by electron microscopy and a concomitant loss of ~50% of the vWF function. © 1982, Rockefeller University Press., All rights reserved.

Duke Scholars

Published In

Journal of Cell Biology

DOI

EISSN

1540-8140

ISSN

0021-9525

Publication Date

November 1, 1982

Volume

95

Issue

2

Start / End Page

632 / 640

Related Subject Headings

  • Developmental Biology
  • 32 Biomedical and clinical sciences
  • 31 Biological sciences
  • 11 Medical and Health Sciences
  • 06 Biological Sciences
 

Citation

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Ohmori, K., Fretto, L. J., Harrison, R. L., Switzer, M. E. P., Erickson, H. P., & McKee, P. A. (1982). Electron microscopy of human factor VIII/von willebrand glycoprotein: Effect of reducing reagents on structure and function. Journal of Cell Biology, 95(2), 632–640. https://doi.org/10.1083/jcb.95.2.632
Ohmori, K., L. J. Fretto, R. L. Harrison, M. E. P. Switzer, H. P. Erickson, and P. A. McKee. “Electron microscopy of human factor VIII/von willebrand glycoprotein: Effect of reducing reagents on structure and function.” Journal of Cell Biology 95, no. 2 (November 1, 1982): 632–40. https://doi.org/10.1083/jcb.95.2.632.
Ohmori K, Fretto LJ, Harrison RL, Switzer MEP, Erickson HP, McKee PA. Electron microscopy of human factor VIII/von willebrand glycoprotein: Effect of reducing reagents on structure and function. Journal of Cell Biology. 1982 Nov 1;95(2):632–40.
Ohmori, K., et al. “Electron microscopy of human factor VIII/von willebrand glycoprotein: Effect of reducing reagents on structure and function.” Journal of Cell Biology, vol. 95, no. 2, Nov. 1982, pp. 632–40. Scopus, doi:10.1083/jcb.95.2.632.
Ohmori K, Fretto LJ, Harrison RL, Switzer MEP, Erickson HP, McKee PA. Electron microscopy of human factor VIII/von willebrand glycoprotein: Effect of reducing reagents on structure and function. Journal of Cell Biology. 1982 Nov 1;95(2):632–640.

Published In

Journal of Cell Biology

DOI

EISSN

1540-8140

ISSN

0021-9525

Publication Date

November 1, 1982

Volume

95

Issue

2

Start / End Page

632 / 640

Related Subject Headings

  • Developmental Biology
  • 32 Biomedical and clinical sciences
  • 31 Biological sciences
  • 11 Medical and Health Sciences
  • 06 Biological Sciences