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. 1985 Jun 1;100(6):1955–1961. doi: 10.1083/jcb.100.6.1955

The binding of vimentin to human erythrocyte membranes: a model system for the study of intermediate filament-membrane interactions

PMCID: PMC2113610  PMID: 3158664

Abstract

We have characterized the association of the intermediate filament protein, vimentin, with the plasma membrane, using radioiodinated lens vimentin and various preparations of human erythrocyte membrane vesicles. Inside-out membrane vesicles (IOVs), depleted of spectrin and actin, bind I125-vimentin in a saturable manner unlike resealed, right- side-out membranes which bind negligible amounts of vimentin in an unsaturable fashion. The binding of vimentin to IOVs is abolished by trypsin or acid treatment of the vesicles. Extraction of protein 4.1 or reconstitution of the membranes with purified spectrin do not basically affect the association. However, removal of ankyrin (band 2.1) significantly lowers the binding. Upon reconstitution of depleted vesicles with purified ankyrin, the vimentin binding function is restored. If ankyrin is added in excess the binding of vimentin to IOVs is quantitatively inhibited, whereas protein 4.1, the cytoplasmic fragment of band 3, band 6, band 4.5 (catalase), or bovine serum albumin do not influence it. Preincubation of the IOVs with a polyclonal anti-ankyrin antibody blocks 90% of the binding. Preimmune sera and antibodies against spectrin, protein 4.1, glycophorin A, and band 3 exhibit no effect. On the basis of these data, we propose that vimentin is able to associate specifically with the erythrocyte membrane skeleton and that ankyrin constitutes its major attachment site.

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Selected References

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  1. Ben-Ze'ev A., Duerr A., Solomon F., Penman S. The outer boundary of the cytoskeleton: a lamina derived from plasma membrane proteins. Cell. 1979 Aug;17(4):859–865. doi: 10.1016/0092-8674(79)90326-x. [DOI] [PubMed] [Google Scholar]
  2. Bennett V., Branton D. Selective association of spectrin with the cytoplasmic surface of human erythrocyte plasma membranes. Quantitative determination with purified (32P)spectrin. J Biol Chem. 1977 Apr 25;252(8):2753–2763. [PubMed] [Google Scholar]
  3. Bennett V., Davis J., Fowler W. E. Brain spectrin, a membrane-associated protein related in structure and function to erythrocyte spectrin. Nature. 1982 Sep 9;299(5879):126–131. doi: 10.1038/299126a0. [DOI] [PubMed] [Google Scholar]
  4. Bennett V., Stenbuck P. J. Association between ankyrin and the cytoplasmic domain of band 3 isolated from the human erythrocyte membrane. J Biol Chem. 1980 Jul 10;255(13):6424–6432. [PubMed] [Google Scholar]
  5. Burridge K., Kelly T., Mangeat P. Nonerythrocyte spectrins: actin-membrane attachment proteins occurring in many cell types. J Cell Biol. 1982 Nov;95(2 Pt 1):478–486. doi: 10.1083/jcb.95.2.478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Capetanaki Y. G., Ngai J., Flytzanis C. N., Lazarides E. Tissue-specific expression of two mRNA species transcribed from a single vimentin gene. Cell. 1983 Dec;35(2 Pt 1):411–420. doi: 10.1016/0092-8674(83)90174-5. [DOI] [PubMed] [Google Scholar]
  7. Cotmore S. F., Furthmayr H., Marchesi V. T. Immunochemical evidence for the transmembrane orientation of glycophorin A. Localization of ferritin-antibody conjugates in intact cells. J Mol Biol. 1977 Jul 5;113(3):539–553. doi: 10.1016/0022-2836(77)90237-6. [DOI] [PubMed] [Google Scholar]
  8. Davis J. Q., Bennett V. Brain ankyrin. A membrane-associated protein with binding sites for spectrin, tubulin, and the cytoplasmic domain of the erythrocyte anion channel. J Biol Chem. 1984 Nov 10;259(21):13550–13559. [PubMed] [Google Scholar]
  9. Davis J. Q., Bennett V. Brain ankyrin. Purification of a 72,000 Mr spectrin-binding domain. J Biol Chem. 1984 Feb 10;259(3):1874–1881. [PubMed] [Google Scholar]
  10. Dellagi K., Vainchenker W., Vinci G., Paulin D., Brouet J. C. Alteration of vimentin intermediate filament expression during differentiation of human hemopoietic cells. EMBO J. 1983;2(9):1509–1514. doi: 10.1002/j.1460-2075.1983.tb01615.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Engel J., Fasold H., Hulla F. W., Waechter F., Wegner A. The polymerization reaction of muscle actin. Mol Cell Biochem. 1977 Nov 25;18(1):3–13. doi: 10.1007/BF00215273. [DOI] [PubMed] [Google Scholar]
  12. Fukuda M., Eshdat Y., Tarone G., Marchesi V. T. Isolation and characterization of peptides derived from the cytoplasmic segment of band 3, the predominant intrinsic membrane protein of the human erythrocyte. J Biol Chem. 1978 Apr 10;253(7):2419–2428. [PubMed] [Google Scholar]
  13. Geisler N., Weber K. The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins. EMBO J. 1982;1(12):1649–1656. doi: 10.1002/j.1460-2075.1982.tb01368.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Glenney J. R., Jr, Glenney P., Weber K. Erythroid spectrin, brain fodrin, and intestinal brush border proteins (TW-260/240) are related molecules containing a common calmodulin-binding subunit bound to a variant cell type-specific subunit. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4002–4005. doi: 10.1073/pnas.79.13.4002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goodman S. R., Zagon I. S., Kulikowski R. R. Identification of a spectrin-like protein in nonerythroid cells. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7570–7574. doi: 10.1073/pnas.78.12.7570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Granger B. L., Lazarides E. Structural associations of synemin and vimentin filaments in avian erythrocytes revealed by immunoelectron microscopy. Cell. 1982 Aug;30(1):263–275. doi: 10.1016/0092-8674(82)90032-0. [DOI] [PubMed] [Google Scholar]
  17. Granger B. L., Repasky E. A., Lazarides E. Synemin and vimentin are components of intermediate filaments in avian erythrocytes. J Cell Biol. 1982 Feb;92(2):299–312. doi: 10.1083/jcb.92.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hargreaves W. R., Giedd K. N., Verkleij A., Branton D. Reassociation of ankyrin with band 3 in erythrocyte membranes and in lipid vesicles. J Biol Chem. 1980 Dec 25;255(24):11965–11972. [PubMed] [Google Scholar]
  19. Kartenbeck J., Schwechheimer K., Moll R., Franke W. W. Attachment of vimentin filaments to desmosomal plaques in human meningiomal cells and arachnoidal tissue. J Cell Biol. 1984 Mar;98(3):1072–1081. doi: 10.1083/jcb.98.3.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Korn E. D. Actin polymerization and its regulation by proteins from nonmuscle cells. Physiol Rev. 1982 Apr;62(2):672–737. doi: 10.1152/physrev.1982.62.2.672. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. 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]
  23. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  24. Lehto V. P., Virtanen I., Kurki P. Intermediate filaments anchor the nuclei in nuclear monolayers of cultured human fibroblasts. Nature. 1978 Mar 9;272(5649):175–177. doi: 10.1038/272175a0. [DOI] [PubMed] [Google Scholar]
  25. Leto T. L., Marchesi V. T. A structural model of human erythrocyte protein 4.1. J Biol Chem. 1984 Apr 10;259(7):4603–4608. [PubMed] [Google Scholar]
  26. Levine J., Willard M. Fodrin: axonally transported polypeptides associated with the internal periphery of many cells. J Cell Biol. 1981 Sep;90(3):631–642. doi: 10.1083/jcb.90.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McNutt N. S., Weinstein R. S. Membrane ultrastructure at mammalian intercellular junctions. Prog Biophys Mol Biol. 1973;26:45–101. doi: 10.1016/0079-6107(73)90017-5. [DOI] [PubMed] [Google Scholar]
  28. Nelson W. J., Traub P. Fractionation of the detergent-resistant filamentous network of Ehrlich ascites tumour cells. Eur J Cell Biol. 1981 Feb;23(2):250–257. [PubMed] [Google Scholar]
  29. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  30. Osborn M., Born T., Koitsch H. J., Weber K. Stereo immunofluorescence microscopy: I. Three-dimensional arrangement of microfilaments, microtubules and tonofilaments. Cell. 1978 Jul;14(3):477–488. doi: 10.1016/0092-8674(78)90234-9. [DOI] [PubMed] [Google Scholar]
  31. Ramaekers F. C., Dunia I., Dodemont H. J., Benedetti E. L., Bloemendal H. Lenticular intermediate-sized filaments: biosynthesis and interaction with plasma membrane. Proc Natl Acad Sci U S A. 1982 May;79(10):3208–3212. doi: 10.1073/pnas.79.10.3208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Repasky E. A., Granger B. L., Lazarides E. Widespread occurrence of avian spectrin in nonerythroid cells. Cell. 1982 Jul;29(3):821–833. doi: 10.1016/0092-8674(82)90444-5. [DOI] [PubMed] [Google Scholar]
  33. Steinert P. M., Idler W. W., Cabral F., Gottesman M. M., Goldman R. D. In vitro assembly of homopolymer and copolymer filaments from intermediate filament subunits of muscle and fibroblastic cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3692–3696. doi: 10.1073/pnas.78.6.3692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weaver D. C., Marchesi V. T. The structural basis of ankyrin function. I. Identification of two structural domains. J Biol Chem. 1984 May 25;259(10):6165–6169. [PubMed] [Google Scholar]
  35. Ziparo E., Lemay A., Marchesi V. T. The distribution of spectrin along the membranes of normal and echinocytic human erythrocytes. J Cell Sci. 1978 Dec;34:91–101. doi: 10.1242/jcs.34.1.91. [DOI] [PubMed] [Google Scholar]

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