Home LiteratureArticle Details
PMID: 3090053 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Vinculin interaction with permeabilized cells: disruption and reconstitution of a binding site.

The Journal of cell biology ·Vol. 103 ·No. 2 ·1986-08-00 ·Pages 641-8

Ball EH, Freitag C, Gurofsky S

Abstract

Fluorescently labeled vinculin binds to focal contact areas in permeabilized cells independent of actin (Avnur, Z., J. V. Small, and B. Geiger, 1983, J. Cell Biol., 96:1622-1630), but the nature of the binding site is unknown. In this study we have examined the interaction of vinculin with these sites in permeabilized L6 myoblasts to define conditions that perturb the binding and subsequently to reconstitute it. Mild treatment with low concentrations of protease prevents vinculin incorporation without gross changes in the cytoskeleton or extensive protein breakdown. Exposure to buffers of moderate ionic strength also reduces subsequent vinculin binding without large morphological effects. These extraction conditions were used to obtain a fraction from gizzard which was able to restore the vinculin localization. Talin, actin, and vinculin itself were able to alter the binding of labeled vinculin to permeabilized cells and each also interacted with vinculin in gel overlays; however, they were unable to reconstitute the binding site in treated permeabilized cells. The results show a requirement for an as yet unidentified protein to capacitate vinculin binding to focal contact sites and suggest that this protein is peripheral and interacts directly with the binding site.

MeSH Terms
Animals Binding Sites Cell Adhesion Cell Line Cell Membrane Permeability Chickens Fluorescent Antibody Technique Gizzard, Avian/ultrastructure Mice Molecular Weight Muscle Proteins/metabolism Muscles/cytology,metabolism Protein Binding Vinculin
Chemicals
Muscle Proteins Vinculin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ball E H
Freitag C
Gurofsky S
References (37)
37 references, click to expand
  1. Retention of differentiation potentialities during prolonged cultivation of myogenic cells.
    Proc Natl Acad Sci U S A. 1968 Oct;61(2):477-83 PMID: 5245982
  2. An integral glycoprotein associated with the membrane attachment sites of actin microfilaments.
    J Cell Biol. 1985 Sep;101(3):785-801 PMID: 3897248
  3. Determination of protein: a modification of the Lowry method that gives a linear photometric response.
    Anal Biochem. 1972 Aug;48(2):422-7 PMID: 4115981
  4. Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril.
    Biochem Biophys Res Commun. 1978 Feb 28;80(4):849-57 PMID: 637870
  5. The outer boundary of the cytoskeleton: a lamina derived from plasma membrane proteins.
    Cell. 1979 Aug;17(4):859-65 PMID: 573666
  6. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells.
    Cell. 1979 Sep;18(1):193-205 PMID: 574428
  7. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes.
    Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127-31 PMID: 6776523
  8. Stress fiber sarcomeres of fibroblasts are contractile.
    Cell. 1980 Nov;22(2 Pt 2):555-61 PMID: 6893813
  9. Altered distributions of the cytoskeletal proteins vinculin and alpha-actinin in cultured fibroblasts transformed by Rous sarcoma virus.
    Proc Natl Acad Sci U S A. 1980 Nov;77(11):6687-91 PMID: 6256755
  10. Vinculin: a cytoskeletal target of the transforming protein of Rous sarcoma virus.
    Cell. 1981 Apr;24(1):165-74 PMID: 6263485
  11. The association of rhodamine - labelled alpha-actinin with actin bundles in demembranated cells.
    Cell Biol Int Rep. 1981 Jun;5(6):627-34 PMID: 6788382
  12. Interaction of alpha-actinin and vinculin with actin: opposite effects on filament network formation.
    Proc Natl Acad Sci U S A. 1981 May;78(5):3005-9 PMID: 6789327
  13. High-affinity interaction of vinculin with actin filaments in vitro.
    Cell. 1982 Jan;28(1):83-90 PMID: 6802502
  14. Involvement of vinculin in contact-induced cytoskeletal interactions.
    Cold Spring Harb Symp Quant Biol. 1982;46 Pt 2:671-82 PMID: 6286219
  15. Intracellular and extracellular components involved in the formation of ventral surfaces of fibroblasts.
    Cold Spring Harb Symp Quant Biol. 1982;46 Pt 2:755-67 PMID: 6809405
  16. Phospholipids stimulate phosphorylation of vinculin by the tyrosine-specific protein kinase of Rous sarcoma virus.
    Proc Natl Acad Sci U S A. 1982 Aug;79(15):4628-31 PMID: 6181509
  17. New surface component of fibroblast's focal contacts identified by a monoclonal antibody.
    Cell. 1982 Dec;31(3 Pt 2):671-9 PMID: 6760990
  18. The spectrin membrane skeleton of normal and abnormal human erythrocytes: a review.
    Am J Physiol. 1983 Mar;244(3):C121-41 PMID: 6338732
  19. Actin-independent association of vinculin with the cytoplasmic aspect of the plasma membrane in cell-contact areas.
    J Cell Biol. 1983 Jun;96(6):1622-30 PMID: 6406516
  20. Membrane-cytoskeleton interaction.
    Biochim Biophys Acta. 1983 Aug 11;737(3-4):305-41 PMID: 6411121
  21. A new protein of adhesion plaques and ruffling membranes.
    J Cell Biol. 1983 Aug;97(2):359-67 PMID: 6684120
  22. Vinculin, a cytoskeletal substrate of protein kinase C.
    J Biol Chem. 1983 Oct 10;258(19):11423-6 PMID: 6225775
  23. Detection of vinculin-binding proteins with an 125I-vinculin gel overlay technique.
    J Cell Biol. 1983 Oct;97(4):1283-7 PMID: 6413514
  24. Vinculin phosphorylation by the src kinase. Interaction of vinculin with phospholipid vesicles.
    J Biol Chem. 1983 Dec 10;258(23):14626-31 PMID: 6417142
  25. Detection of high molecular weight vinculin binding proteins in muscle and nonmuscle tissues with an electroblot-overlay technique.
    Biochem Biophys Res Commun. 1983 Nov 15;116(3):1026-32 PMID: 6418169
  26. Talin: a cytoskeletal component concentrated in adhesion plaques and other sites of actin-membrane interaction.
    Cell Motil. 1983;3(5-6):405-17 PMID: 6319001
  27. A 200-kd protein isolated from the fascia adherens membrane domains of chicken cardiac muscle cells is detected immunologically in fibroblast focal adhesions.
    Cell Motil. 1983;3(5-6):419-29 PMID: 6362888
  28. Properties of smooth muscle vinculin.
    J Biol Chem. 1984 Mar 25;259(6):3916-24 PMID: 6323447
  29. Vinculin phosphorylation in response to calcium and phorbol esters in intact cells.
    J Biol Chem. 1984 Apr 25;259(8):5264-70 PMID: 6325428
  30. An interaction between vinculin and talin.
    Nature. 1984 Apr 19-25;308(5961):744-6 PMID: 6425696
  31. Actin-membrane interaction in fibroblasts: what proteins are involved in this association?
    J Cell Biol. 1984 Jul;99(1 Pt 2):95s-103s PMID: 6430913
  32. A tumor promoter induces rapid and coordinated reorganization of actin and vinculin in cultured cells.
    J Cell Biol. 1984 Sep;99(3):1045-59 PMID: 6206076
  33. Interaction of fluorescently-labeled contractile proteins with the cytoskeleton in cell models.
    J Cell Biol. 1984 Sep;99(3):918-28 PMID: 6540785
  34. Smooth muscle caldesmon. Rapid purification and F-actin cross-linking properties.
    J Biol Chem. 1984 Oct 25;259(20):12873-80 PMID: 6092349
  35. Isolation and partial characterization of human platelet vinculin.
    J Cell Biol. 1985 Mar;100(3):669-76 PMID: 3919032
  36. Focal contacts of normal and RSV-transformed quail cells. Hypothesis of the transformation-induced deficient maturation of focal contacts.
    Exp Cell Res. 1985 Jun;158(2):433-44 PMID: 2988988
  37. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.103.2.641
Published
1986-08-00
Pages
641-8
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113843
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.103.2.641 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2