Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1993 Jun 1; 121(5): 1065–1073.
PMCID: PMC2119678

Purification and properties of transgelin: a transformation and shape change sensitive actin-gelling protein


We have purified the transformation and shape change sensitive isoform of an actin associated polypeptide doublet previously described by us (Shapland, C., P. Lowings, and D. Lawson. 1988. J. Cell Biol. 107:153- 161) and have shown that it is evolutionarily conserved as far back as yeast. The purified protein: (a) binds directly to actin filaments at a ratio of 1:6 actin monomers, with a binding constant (Ka) of approximately 7.5 x 10(5) M-1; and (b) causes actin filament gelation within 2 min. Although these activities are controlled by ionic strength (and may be mediated by positively charged amino acid residues) the molecule remains as a monomer irrespective of ionic conditions. EM reveals that the addition of this protein to actin filaments converts them from a loose, random distribution into a tangled, cross-linked meshwork within 1 min, and discrete tightly aggregated foci after 10 min. By use of an "add-back" cell permeabilization system we can rebind this molecule specifically to actin filaments in cells from which it has previously been removed. Since the protein is transformation sensitive and gels actin, we have named it transgelin.

Full Text

The Full Text of this article is available as a PDF (1.9M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Almendral JM, Santarén JF, Perera J, Zerial M, Bravo R. Expression, cloning and cDNA sequence of a fibroblast serum-regulated gene encoding a putative actin-associated protein (p27). Exp Cell Res. 1989 Apr;181(2):518–530. [PubMed]
  • Bretscher A. Fimbrin is a cytoskeletal protein that crosslinks F-actin in vitro. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6849–6853. [PMC free article] [PubMed]
  • de Arruda MV, Watson S, Lin CS, Leavitt J, Matsudaira P. Fimbrin is a homologue of the cytoplasmic phosphoprotein plastin and has domains homologous with calmodulin and actin gelation proteins. J Cell Biol. 1990 Sep;111(3):1069–1079. [PMC free article] [PubMed]
  • Drenckhahn D, Engel K, Höfer D, Merte C, Tilney L, Tilney M. Three different actin filament assemblies occur in every hair cell: each contains a specific actin crosslinking protein. J Cell Biol. 1991 Feb;112(4):641–651. [PMC free article] [PubMed]
  • Griffith LM, Pollard TD. Cross-linking of actin filament networks by self-association and actin-binding macromolecules. J Biol Chem. 1982 Aug 10;257(15):9135–9142. [PubMed]
  • Hartwig JH, DeSisto M. The cytoskeleton of the resting human blood platelet: structure of the membrane skeleton and its attachment to actin filaments. J Cell Biol. 1991 Feb;112(3):407–425. [PMC free article] [PubMed]
  • Hartwig JH, Kwiatkowski DJ. Actin-binding proteins. Curr Opin Cell Biol. 1991 Feb;3(1):87–97. [PubMed]
  • Hartwig JH, Stossel TP. Isolation and properties of actin, myosin, and a new actinbinding protein in rabbit alveolar macrophages. J Biol Chem. 1975 Jul 25;250(14):5696–5705. [PubMed]
  • Hendricks M, Weintraub H. Tropomyosin is decreased in transformed cells. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5633–5637. [PMC free article] [PubMed]
  • Husain-Chishti A, Faquin W, Wu CC, Branton D. Purification of erythrocyte dematin (protein 4.9) reveals an endogenous protein kinase that modulates actin-bundling activity. J Biol Chem. 1989 May 25;264(15):8985–8991. [PubMed]
  • Koffer A, Gratzer WB, Clarke GD, Hales A. Phase equilibria of cytoplasmic actin of cultured epithelial (BHK) cells. J Cell Sci. 1983 May;61:191–218. [PubMed]
  • Owada MK, Hakura A, Iida K, Yahara I, Sobue K, Kakiuchi S. Occurrence of caldesmon (a calmodulin-binding protein) in cultured cells: comparison of normal and transformed cells. Proc Natl Acad Sci U S A. 1984 May;81(10):3133–3137. [PMC free article] [PubMed]
  • Korn ED. Actin polymerization and its regulation by proteins from nonmuscle cells. Physiol Rev. 1982 Apr;62(2):672–737. [PubMed]
  • Kwiatkowski DJ. Predominant induction of gelsolin and actin-binding protein during myeloid differentiation. J Biol Chem. 1988 Sep 25;263(27):13857–13862. [PubMed]
  • Lawson D. Epinemin: a new protein associated with vimentin filaments in non-neural cells. J Cell Biol. 1983 Dec;97(6):1891–1905. [PMC free article] [PubMed]
  • Lawson D. Distribution of myosin and relationship to actin organization in cortical and subcortical areas of antibody-labelled, quick-frozen, deep-etched fibroblast cytoskeletons. Cell Motil Cytoskeleton. 1987;7(4):368–380. [PubMed]
  • Lazarides E. Actin, alpha-actinin, and tropomyosin interaction in the structural organization of actin filaments in nonmuscle cells. J Cell Biol. 1976 Feb;68(2):202–219. [PMC free article] [PubMed]
  • Leavitt J, Gunning P, Kedes L, Jariwalla R. Smooth muscle alpha-action is a transformation-sensitive marker for mouse NIH 3T3 and Rat-2 cells. Nature. 316(6031):840–842. [PubMed]
  • Lees-Miller JP, Heeley DH, Smillie LB. An abundant and novel protein of 22 kDa (SM22) is widely distributed in smooth muscles. Purification from bovine aorta. Biochem J. 1987 Jun 15;244(3):705–709. [PMC free article] [PubMed]
  • Lees-Miller JP, Heeley DH, Smillie LB, Kay CM. Isolation and characterization of an abundant and novel 22-kDa protein (SM22) from chicken gizzard smooth muscle. J Biol Chem. 1987 Mar 5;262(7):2988–2993. [PubMed]
  • MacLean-Fletcher SD, Pollard TD. Viscometric analysis of the gelation of Acanthamoeba extracts and purification of two gelation factors. J Cell Biol. 1980 May;85(2):414–428. [PMC free article] [PubMed]
  • MARTIN RG, AMES BN. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed]
  • Maruta H, Korn ED. Purification from Acanthamoeba castellanii of proteins that induce gelation and syneresis of F-actin. J Biol Chem. 1977 Jan 10;252(1):399–402. [PubMed]
  • Matsudaira P. Modular organization of actin crosslinking proteins. Trends Biochem Sci. 1991 Mar;16(3):87–92. [PubMed]
  • Muhlrad A. The isolated 21 kDa N-terminal fragment of myosin binds to actin in an ATP and ionic strength-dependent manner. Biochim Biophys Acta. 1991 Apr 29;1077(3):308–315. [PubMed]
  • Murphy DB, Gray RO, Grasser WA, Pollard TD. Direct demonstration of actin filament annealing in vitro. J Cell Biol. 1988 Jun;106(6):1947–1954. [PMC free article] [PubMed]
  • Nishida W, Kitami Y, Abe M, Hiwada K. Gene cloning and nucleotide sequence of SM22 alpha from the chicken gizzard smooth muscle. Biochem Int. 1991 Mar;23(4):663–668. [PubMed]
  • Pardee JD, Spudich JA. Purification of muscle actin. Methods Enzymol. 1982;85(Pt B):164–181. [PubMed]
  • Pagliaro L, Taylor DL. Aldolase exists in both the fluid and solid phases of cytoplasm. J Cell Biol. 1988 Sep;107(3):981–991. [PMC free article] [PubMed]
  • Pearlstone JR, Weber M, Lees-Miller JP, Carpenter MR, Smillie LB. Amino acid sequence of chicken gizzard smooth muscle SM22 alpha. J Biol Chem. 1987 May 5;262(13):5985–5991. [PubMed]
  • Pollard TD. Cytoplasmic contractile proteins. J Cell Biol. 1981 Dec;91(3 Pt 2):156s–165s. [PMC free article] [PubMed]
  • Pollard TD. Actin. Curr Opin Cell Biol. 1990 Feb;2(1):33–40. [PubMed]
  • Pollard TD, Cooper JA. Methods to characterize actin filament networks. Methods Enzymol. 1982;85(Pt B):211–233. [PubMed]
  • Pollard TD, Cooper JA. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. [PubMed]
  • Santarén JF, Blüthmann H, MacDonald-Bravo H, Bravo R. Specific antibody against a protein (p27) present in nonestablished fibroblasts. A putative microfilament-associated protein. Exp Cell Res. 1987 Dec;173(2):341–348. [PubMed]
  • Schleicher M, André E, Hartmann H, Noegel AA. Actin-binding proteins are conserved from slime molds to man. Dev Genet. 1988;9(4-5):521–530. [PubMed]
  • Schliwa M. Proteins associated with cytoplasmic actin. Cell. 1981 Sep;25(3):587–590. [PubMed]
  • Shapland C, Lowings P, Lawson D. Identification of new actin-associated polypeptides that are modified by viral transformation and changes in cell shape. J Cell Biol. 1988 Jul;107(1):153–161. [PMC free article] [PubMed]
  • Spudich JA, Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed]
  • Stossel TP. From signal to pseudopod. How cells control cytoplasmic actin assembly. J Biol Chem. 1989 Nov 5;264(31):18261–18264. [PubMed]
  • Stossel TP, Chaponnier C, Ezzell RM, Hartwig JH, Janmey PA, Kwiatkowski DJ, Lind SE, Smith DB, Southwick FS, Yin HL, et al. Nonmuscle actin-binding proteins. Annu Rev Cell Biol. 1985;1:353–402. [PubMed]
  • Thweatt R, Lumpkin CK, Jr, Goldstein S. A novel gene encoding a smooth muscle protein is overexpressed in senescent human fibroblasts. Biochem Biophys Res Commun. 1992 Aug 31;187(1):1–7. [PubMed]
  • Tilney LG. Actin filaments in the acrosomal reaction of Limulus sperm. Motion generated by alterations in the packing of the filaments. J Cell Biol. 1975 Feb;64(2):289–310. [PMC free article] [PubMed]
  • Totty NF, Waterfield MD, Hsuan JJ. Accelerated high-sensitivity microsequencing of proteins and peptides using a miniature reaction cartridge. Protein Sci. 1992 Sep;1(9):1215–1224. [PMC free article] [PubMed]
  • Vancompernolle K, Gimona M, Herzog M, Van Damme J, Vandekerckhove J, Small V. Isolation and sequence of a tropomyosin-binding fragment of turkey gizzard calponin. FEBS Lett. 1990 Nov 12;274(1-2):146–150. [PubMed]
  • Vandekerckhove J. Actin-binding proteins. Curr Opin Cell Biol. 1990 Feb;2(1):41–50. [PubMed]
  • Vandekerckhove J, Bauw G, Vancompernolle K, Honoré B, Celis J. Comparative two-dimensional gel analysis and microsequencing identifies gelsolin as one of the most prominent downregulated markers of transformed human fibroblast and epithelial cells. J Cell Biol. 1990 Jul;111(1):95–102. [PMC free article] [PubMed]
  • Van Roy F, Mareel M. Tumour invasion: effects of cell adhesion and motility. Trends Cell Biol. 1992 Jun;2(6):163–169. [PubMed]
  • Walsh MP. The Ayerst Award Lecture 1990. Calcium-dependent mechanisms of regulation of smooth muscle contraction. Biochem Cell Biol. 1991 Dec;69(12):771–800. [PubMed]
  • Wang K. Filamin, a new high-molecular-weight protein found in smooth muscle and nonmuscle cells. Purification and properties of chicken gizzard filamin. Biochemistry. 1977 May 3;16(9):1857–1865. [PubMed]
  • Wang K, Feramisco JR, Ash JF. Fluorescent localization of contractile proteins in tissue culture cells. Methods Enzymol. 1982;85(Pt B):514–562. [PubMed]
  • Way M, Weeds A. Actin-binding proteins. Cytoskeletal ups and downs. Nature. 1990 Mar 22;344(6264):292–294. [PubMed]
  • Weeds A. Actin-binding proteins--regulators of cell architecture and motility. Nature. 1982 Apr 29;296(5860):811–816. [PubMed]
  • Wilkins JA, Lin S. A re-examination of the interaction of vinculin with actin. J Cell Biol. 1986 Mar;102(3):1085–1092. [PMC free article] [PubMed]
  • Yamashiro-Matsumura S, Matsumura F. Characterization of 83-kilodalton nonmuscle caldesmon from cultured rat cells: stimulation of actin binding of nonmuscle tropomyosin and periodic localization along microfilaments like tropomyosin. J Cell Biol. 1988 Jun;106(6):1973–1983. [PMC free article] [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • OMIM
    Genome Survey Sequence (GSS) nucleotide records reported in the current articles.
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...