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PMID: 8161679 Published · ppublish English Journal Article

Three-dimensional reconstruction of a co-complex of F-actin with antibody Fab fragments to actin's NH2 terminus.

Biophysical journal ·Vol. 66 ·No. 2 Pt 1 ·1994-02-00 ·Pages 276-85

Orlova A, Yu X, Egelman EH

Abstract

We have decorated F-actin with Fab fragments of antibodies to actin residues 1-7. These antibody fragments do not strongly affect the rigor binding of myosin S-1 to actin, but do affect the binding of S-1 to actin in the presence of nucleotide (DasGupta, G., and E. Reisler, 1989. J. Mol. Biol. 207:833-836; 1991. Biochemistry. 30:9961-9966; 1992. Biochemistry. 31:1836-1841). Although the binding constant is rather low, we estimate that we have achieved about 85% occupancy of the actin sites. Three-dimensional reconstructions from electron micrographs of both negatively stained and frozen-hydrated filaments show that the Fab fragment is bound at the location of the NH2 terminus in the model of Holmes et al. (Holmes, K.C., D. Popp, W. Gebhard, and W. Kabsch. 1990. Nature. 347:37-44) for F-actin, excluding very different orientations of the actin subunit in the filament. Most of the mass of the antibody is not visualized, which is due to the large mobility of the NH2 terminus in F-actin, differences in binding angle within the polyclonal antibody population, or a combination of both of these possibilities.

MeSH Terms
Actins/chemistry,immunology,ultrastructure Adenosine Triphosphate/chemistry Animals Antigen-Antibody Complex/chemistry,ultrastructure Binding Sites Biophysical Phenomena Biophysics Erythrosine Image Processing, Computer-Assisted Immunoglobulin Fab Fragments/chemistry Microscopy, Electron Models, Molecular Myosin Subfragments/chemistry Phalloidine Protein Binding Protein Conformation Rabbits
Chemicals
Actins Antigen-Antibody Complex Immunoglobulin Fab Fragments Myosin Subfragments Phalloidine Adenosine Triphosphate Erythrosine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Orlova A
Department of Cell Biology and Neuroanatomy, University of Minnesota Medical School, Minneapolis 55455.
Yu X
Egelman E H
References (35)
35 references, click to expand
  1. Electron microscopy of thin filaments decorated with a Ca2+-regulated myosin.
    J Mol Biol. 1980 Jun 15;140(1):35-55 PMID: 6997502
  2. Electron microscopic evidence for the axial rotation and inter-domain flexibility of the Fab regions of immunoglobulin G.
    J Mol Biol. 1983 Sep 25;169(3):771-4 PMID: 6631952
  3. Identification of myosin-binding sites on the actin sequence.
    Biochemistry. 1982 Jul 20;21(15):3654-61 PMID: 7115691
  4. Peptide antibody specific for the amino terminus of skeletal muscle alpha-actin.
    Proc Natl Acad Sci U S A. 1983 Mar;80(6):1506-10 PMID: 6572911
  5. Stoichiometry of actin X S-1 cross-linked complex.
    J Biol Chem. 1984 Jun 25;259(12):7363-6 PMID: 6736009
  6. Cross-linking of actin to myosin subfragment 1: course of reaction and stoichiometry of products.
    Biochemistry. 1985 Jan 1;24(1):137-44 PMID: 3846455
  7. An algorithm for straightening images of curved filamentous structures.
    Ultramicroscopy. 1986;19(4):367-73 PMID: 3775966
  8. Structural relationships of actin, myosin, and tropomyosin revealed by cryo-electron microscopy.
    J Cell Biol. 1987 Jul;105(1):29-39 PMID: 3611188
  9. Antibodies directed against N-terminal residues on actin do not block acto-myosin binding.
    Biochemistry. 1987 Sep 22;26(19):6064-70 PMID: 3689759
  10. The location of DNA in RecA-DNA helical filaments.
    Science. 1989 Jul 28;245(4916):404-7 PMID: 2667137
  11. Antibody against the amino terminus of alpha-actin inhibits actomyosin interactions in the presence of ATP.
    J Mol Biol. 1989 Jun 20;207(4):833-6 PMID: 2760933
  12. Localization of VP4 neutralization sites in rotavirus by three-dimensional cryo-electron microscopy.
    Nature. 1990 Feb 1;343(6257):476-9 PMID: 2153941
  13. Atomic structure of the actin:DNase I complex.
    Nature. 1990 Sep 6;347(6288):37-44 PMID: 2395459
  14. Atomic model of the actin filament.
    Nature. 1990 Sep 6;347(6288):44-9 PMID: 2395461
  15. Interference with myosin subfragment-1 binding by site-directed mutagenesis of actin.
    Eur J Biochem. 1991 Aug 15;200(1):35-41 PMID: 1879430
  16. Site-directed mutations of Dictyostelium actin: disruption of a negative charge cluster at the N terminus.
    Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7711-4 PMID: 1831905
  17. Nucleotide-induced changes in the interaction of myosin subfragment 1 with actin: detection by antibodies against the N-terminal segment of actin.
    Biochemistry. 1991 Oct 15;30(41):9961-6 PMID: 1911787
  18. The structural basis for the intrinsic disorder of the actin filament: the "lateral slipping" model.
    J Cell Biol. 1991 Nov;115(3):689-703 PMID: 1918159
  19. Identification of a Fab interaction footprint site on an icosahedral virus by cryoelectron microscopy and X-ray crystallography.
    Nature. 1992 Jan 16;355(6357):275-8 PMID: 1731227
  20. Actomyosin interactions in the presence of ATP and the N-terminal segment of actin.
    Biochemistry. 1992 Feb 18;31(6):1836-41 PMID: 1531299
  21. Removal of the amino-terminal acidic residues of yeast actin. Studies in vitro and in vivo.
    J Biol Chem. 1992 May 5;267(13):9430-6 PMID: 1349604
  22. 3D reconstruction from the Fourier transform of a single superlattice image of an oblique section.
    Ultramicroscopy. 1992 Apr-May;41(1-3):153-67 PMID: 1641913
  23. Effect of trifluoroethanol on protein secondary structure: an NMR and CD study using a synthetic actin peptide.
    Biochemistry. 1992 Sep 22;31(37):8790-8 PMID: 1390666
  24. Structural basis for the destabilization of F-actin by phosphate release following ATP hydrolysis.
    J Mol Biol. 1992 Oct 20;227(4):1043-53 PMID: 1433285
  25. The three-dimensional structure of an intact monoclonal antibody for canine lymphoma.
    Nature. 1992 Nov 26;360(6402):369-72 PMID: 1448155
  26. Image analysis shows that variations in actin crossover spacings are random, not compensatory.
    Biophys J. 1992 Nov;63(5):1299-305 PMID: 1477281
  27. Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus.
    J Biol Chem. 1993 Feb 5;268(4):2410-5 PMID: 8428914
  28. Structure of human rhinovirus complexed with Fab fragments from a neutralizing antibody.
    J Virol. 1993 Mar;67(3):1148-58 PMID: 7679742
  29. Actin isoforms.
    Curr Opin Cell Biol. 1993 Feb;5(1):48-55 PMID: 8448030
  30. Structure of the actin-myosin complex and its implications for muscle contraction.
    Science. 1993 Jul 2;261(5117):58-65 PMID: 8316858
  31. A conformational change in the actin subunit can change the flexibility of the actin filament.
    J Mol Biol. 1993 Jul 20;232(2):334-41 PMID: 8345515
  32. The structure of crystalline profilin-beta-actin.
    Nature. 1993 Oct 28;365(6449):810-6 PMID: 8413665
  33. The variable twist of actin and its modulation by actin-binding proteins.
    J Cell Biol. 1987 Apr;104(4):1005-17 PMID: 3558475
  34. Mapping of actin-binding sites on the heavy chain of myosin subfragment 1.
    Biochemistry. 1983 Mar 29;22(7):1579-85 PMID: 6849869
  35. F-actin is a helix with a random variable twist.
    Nature. 1982 Jul 8;298(5870):131-5 PMID: 7201078
Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/s0006-3495(94)80791-x
Published
1994-02-00
Pages
276-85
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1275692
Subset
IM
External Links
PubMed source DOI: 10.1016/s0006-3495(94)80791-x PMC Full Text View journal details
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