Abstract
Certain smooth muscle cells of blood vessel walls as well as cultured baby hamster kidney cells contain simultaneously two different intermediate-sized filament (IF) proteins, desmin and vimentin. We have examined the question of the occurrence of both proteins in the same IF by chemically crosslinking the single cysteine group present in each of them. Oxidative crosslinking of filaments present in cytoskeletal preparations with cupric ion complexes of 1,10-phenanthroline resulted in formation of three types of dimers: vimentin-vimentin, desmin-desmin, and vimentin-desmin. These dimers were separated by NaDodSO4/polyacrylamide gel electrophoresis and characterized by binding of specific antibodies, by one- and two-dimensional gel electrophoresis of monomers obtained after cleavage of the disulfide bond by thiol agents, and by mapping of radioiodinated tryptic peptides. The demonstration of heterodimers of vimentin and desmin in vascular smooth muscle tissue of cow and chicken and in baby hamster kidney cells shows that the two proteins can be integrated in the same IF and can be nearest neighbors, oriented with their cysteine residues in a mirror-image symmetry. The possible existence of heteropolymer IF in other cell types is discussed.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bennett G. S., Fellini S. A., Croop J. M., Otto J. J., Bryan J., Holtzer H. Differences among 100-A filamentilament subunits from different cell types. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4364–4368. doi: 10.1073/pnas.75.9.4364. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett G. S., Fellini S. A., Holtzer H. Immunofluorescent visualization of 100 A filaments in different cultured chick embryo cell types. Differentiation. 1978;12(2):71–82. doi: 10.1111/j.1432-0436.1979.tb00992.x. [DOI] [PubMed] [Google Scholar]
- Bennett G. S., Fellini S. A., Toyama Y., Holtzer H. Redistribution of intermediate filament subunits during skeletal myogenesis and maturation in vitro. J Cell Biol. 1979 Aug;82(2):577–584. doi: 10.1083/jcb.82.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bragg P. D., Hou C. Subunit composition, function, and spatial arrangement in the Ca2+-and Mg2+-activated adenosine triphosphatases of Escherichia coli and Salmonella typhimurium. Arch Biochem Biophys. 1975 Mar;167(1):311–321. doi: 10.1016/0003-9861(75)90467-1. [DOI] [PubMed] [Google Scholar]
- Bretscher A., Weber K. Villin is a major protein of the microvillus cytoskeleton which binds both G and F actin in a calcium-dependent manner. Cell. 1980 Jul;20(3):839–847. doi: 10.1016/0092-8674(80)90330-x. [DOI] [PubMed] [Google Scholar]
- Cabral F., Gottesman M. M. Phosphorylation of the 10-nm filament protein from Chinese hamster ovary cells. J Biol Chem. 1979 Jul 25;254(14):6203–6206. [PubMed] [Google Scholar]
- Elder J. H., Pickett R. A., 2nd, Hampton J., Lerner R. A. Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. J Biol Chem. 1977 Sep 25;252(18):6510–6515. [PubMed] [Google Scholar]
- Frank E. D., Warren L. Aortic smooth muscle cells contain vimentin instead of desmin. Proc Natl Acad Sci U S A. 1981 May;78(5):3020–3024. doi: 10.1073/pnas.78.5.3020. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Freudenstein C., Appelhans B., Osborn M., Weber K., Keenan T. W. Intermediate-sized filaments of the prekeratin type in myoepithelial cells. J Cell Biol. 1980 Mar;84(3):633–654. doi: 10.1083/jcb.84.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Osborn M., Weber K. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5034–5038. doi: 10.1073/pnas.75.10.5034. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Weber K., Osborn M. HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979 Jan;118(1):95–109. doi: 10.1016/0014-4827(79)90587-1. [DOI] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Winter S., Osborn M., Weber K. Widespread occurrence of intermediate-sized filaments of the vimentin-type in cultured cells from diverse vertebrates. Exp Cell Res. 1979 Oct 1;123(1):25–46. doi: 10.1016/0014-4827(79)90418-x. [DOI] [PubMed] [Google Scholar]
- Gabbiani G., Schmid E., Winter S., Chaponnier C., de Ckhastonay C., Vandekerckhove J., Weber K., Franke W. W. Vascular smooth muscle cells differ from other smooth muscle cells: predominance of vimentin filaments and a specific alpha-type actin. Proc Natl Acad Sci U S A. 1981 Jan;78(1):298–302. doi: 10.1073/pnas.78.1.298. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gard D. L., Bell P. B., Lazarides E. Coexistence of desmin and the fibroblastic intermediate filament subunit in muscle and nonmuscle cells: identification and comparative peptide analysis. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3894–3898. doi: 10.1073/pnas.76.8.3894. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gard D. L., Lazarides E. The synthesis and distribution of desmin and vimentin during myogenesis in vitro. Cell. 1980 Jan;19(1):263–275. doi: 10.1016/0092-8674(80)90408-0. [DOI] [PubMed] [Google Scholar]
- Geisler N., Weber K. Comparison of the proteins of two immunologically distinct intermediate-sized filaments by amino acid sequence analysis: desmin and vimentin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4120–4123. doi: 10.1073/pnas.78.7.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geisler N., Weber K. Self-assembly in Vitro of the 68,000 molecular weight component of the mammalian neurofilament triplet proteins into intermediate-sized filaments. J Mol Biol. 1981 Sep 25;151(3):565–571. doi: 10.1016/0022-2836(81)90011-5. [DOI] [PubMed] [Google Scholar]
- Kobashi K. Catalytic oxidation of sulfhydryl groups by o-phenanthroline copper complex. Biochim Biophys Acta. 1968 May;158(2):239–245. doi: 10.1016/0304-4165(68)90136-0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- O'Connor C. M., Balzer D. R., Jr, Lazarides E. Phosphorylation of subunit proteins of intermediate filaments from chicken muscle and nonmuscle cells. Proc Natl Acad Sci U S A. 1979 Feb;76(2):819–823. doi: 10.1073/pnas.76.2.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Osborn M., Caselitz J., Weber K. Heterogeneity of intermediate filament expression in vascular smooth muscle: a gradient in desmin positive cells from the rat aortic arch to the level of the arteria iliaca communis. Differentiation. 1981;20(3):196–202. doi: 10.1111/j.1432-0436.1981.tb01176.x. [DOI] [PubMed] [Google Scholar]
- Osborn M., Franke W., Weber K. Direct demonstration of the presence of two immunologically distinct intermediate-sized filament systems in the same cell by double immunofluorescence microscopy. Vimentin and cytokeratin fibers in cultured epithelial cells. Exp Cell Res. 1980 Jan;125(1):37–46. doi: 10.1016/0014-4827(80)90186-x. [DOI] [PubMed] [Google Scholar]
- Osborn M., Ludwig-Festl M., Weber K., Bignami A., Dahl D., Bayreuther K. Expression of glial and vimentin type intermediate filaments in cultures derived from human glial material. Differentiation. 1981;19(3):161–167. doi: 10.1111/j.1432-0436.1981.tb01143.x. [DOI] [PubMed] [Google Scholar]
- Paetau A., Virtanen I., Stenman S., Kurki P., Linder E., Vaheri A., Westermark B., Dahl D., Haltia M. Glial fibrillary acidic protein and intermediate filaments in human glioma cells. Acta Neuropathol. 1979 Jun 15;47(1):71–74. doi: 10.1007/BF00698276. [DOI] [PubMed] [Google Scholar]
- Peters K., Richards F. M. Chemical cross-linking: reagents and problems in studies of membrane structure. Annu Rev Biochem. 1977;46:523–551. doi: 10.1146/annurev.bi.46.070177.002515. [DOI] [PubMed] [Google Scholar]
- Renner W., Franke W. W., Schmid E., Geisler N., Weber K., Mandelkow E. Reconstitution of intermediate-sized filaments from denatured monomeric vimentin. J Mol Biol. 1981 Jun 25;149(2):285–306. doi: 10.1016/0022-2836(81)90303-x. [DOI] [PubMed] [Google Scholar]
- Rueger D. C., Huston J. S., Dahl D., Bignami A. Formation of 100 A filaments from purified glial fibrillary acidic protein in vitro. J Mol Biol. 1979 Nov 25;135(1):53–68. doi: 10.1016/0022-2836(79)90340-1. [DOI] [PubMed] [Google Scholar]
- Schmid E., Osborn M., Rungger-Brändle E., Gabbiani G., Weber K., Franke W. W. Distribution of vimentin and desmin filaments in smooth muscle tissue of mammalian and avian aorta. Exp Cell Res. 1982 Feb;137(2):329–340. doi: 10.1016/0014-4827(82)90034-9. [DOI] [PubMed] [Google Scholar]
- Schmid E., Tapscott S., Bennett G. S., Croop J., Fellini S. A., Holtzer H., Franke W. W. Differential location of different types of intermediate-sized filaments in various tissues of the chicken embryo. Differentiation. 1979;15(1):27–40. doi: 10.1111/j.1432-0436.1979.tb01031.x. [DOI] [PubMed] [Google Scholar]
- Schnitzer J., Franke W. W., Schachner M. Immunocytochemical demonstration of vimentin in astrocytes and ependymal cells of developing and adult mouse nervous system. J Cell Biol. 1981 Aug;90(2):435–447. doi: 10.1083/jcb.90.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Steinert P. M., Idler W. W., Goldman R. D. Intermediate filaments of baby hamster kidney (BHK-21) cells and bovine epidermal keratinocytes have similar ultrastructures and subunit domain structures. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4534–4538. doi: 10.1073/pnas.77.8.4534. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinert P. M., Idler W. W., Zimmerman S. B. Self-assembly of bovine epidermal keratin filaments in vitro. J Mol Biol. 1976 Dec 15;108(3):547–567. doi: 10.1016/s0022-2836(76)80136-2. [DOI] [PubMed] [Google Scholar]
- Takahashi M., Takano M., Asada K. Tris-induced cross-linking of thylakoid peptides; thiol oxidation catalyzed by Tris-Cu2+ complexes as a possible mechanism. J Biochem. 1981 Jul;90(1):87–94. doi: 10.1093/oxfordjournals.jbchem.a133472. [DOI] [PubMed] [Google Scholar]
- Tapscott S. J., Bennett G. S., Toyama Y., Kleinbart F., Holtzer H. Intermediate filament proteins in the developing chick spinal cord. Dev Biol. 1981 Aug;86(1):40–54. doi: 10.1016/0012-1606(81)90313-4. [DOI] [PubMed] [Google Scholar]
- Thornton J. M. Disulphide bridges in globular proteins. J Mol Biol. 1981 Sep 15;151(2):261–287. doi: 10.1016/0022-2836(81)90515-5. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tuszynski G. P., Frank E. D., Damsky C. H., Buck C. A., Warren L. The detection of smooth muscle desmin-like protein in BHK21/C13 fibroblasts. J Biol Chem. 1979 Jul 10;254(13):6138–6143. [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
- Yen S. H., Fields K. L. Antibodies to neurofilament, glial filament, and fibroblast intermediate filament proteins bind to different cell types of the nervous system. J Cell Biol. 1981 Jan;88(1):115–126. doi: 10.1083/jcb.88.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zackroff R. V., Goldman R. D. In vitro assembly of intermediate filaments from baby hamster kidney (BHK-21) cells. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6226–6230. doi: 10.1073/pnas.76.12.6226. [DOI] [PMC free article] [PubMed] [Google Scholar]