Home LiteratureArticle Details
PMID: 6195188 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Studies on the antigenic sites of actin: a comparative study of the immunogenic crossreactivity of invertebrate actins.

Journal of muscle research and cell motility ·Vol. 4 ·No. 4 ·1983-08-00 ·Pages 405-27

De Couet HG

Abstract

The structural homologies of invertebrate actins with cytoplasmic vertebrate actins have recently been substantiated by comparative sequence analyses. This suggests that cytoplasmic actin is the ancestral precursor of smooth and striated muscle actin in vertebrates. We have raised antibodies in rabbits against a number of invertebrate muscle actins and have characterized the antisera by means of the highly sensitive ELISA method, which allows quantitation of nanomolar amounts of actin. Despite the fact that the invertebrate actins examined are very similar in primary structure, our results indicate that antibodies raised against them clearly distinguish between only a few amino-acid substitutions, and that the immunoreactivities quantitatively reflect the genetic divergence of this ubiquitous conservative protein. Examination of several proteolytic fragments of scallop actin for immunoreactivity with the homologous antiserum suggests that the major antigenic sites of actin are located within the amino terminal region of the molecule, while a carboxy terminal fragment comprising residues 69-372 exhibits very weak crossreactivity. Immunoadsorption experiments further indicate that species-specific antibodies are directed to antigenic determinants in the N-terminal region. This finding is supported by an examination of the effects of chemical modifications to Tyr, His, Arg, and Cys residues on the immunoreactivity of actin. Interaction with DNAase I markedly decreases the immunoreactivity of actin. This is consonant with the finding that the amino terminal peptide comprising residues 1-207 inhibits DNAase I, whilst a tryptic fragment fails to bind to the enzyme. The interaction is abolished by EDTA and the removal of the tightly bound cation is accompanied by a conformational change, shown by shifts in circular dichroic spectra. The possible involvement of the amino terminal peptide of actin in cation binding is discussed.

MeSH Terms
Actins/immunology Amino Acids/analysis Animals Antibodies/immunology Binding Sites Cross Reactions Deoxyribonuclease I Endodeoxyribonucleases/metabolism Enzyme-Linked Immunosorbent Assay Epitopes/analysis Invertebrates Peptide Fragments/immunology
Chemicals
Actins Amino Acids Antibodies Epitopes Peptide Fragments Endodeoxyribonucleases Deoxyribonuclease I
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
De Couet H G
References (57)
57 references, click to expand
  1. A study of invertebrate actins by isoelectric focusing and immunodiffusion.
    Experientia. 1980 Apr 15;36(4):404-5 PMID: 6769692
  2. Immunological differences between actins from cardiac muscle, skeletal muscle, and brain.
    Proc Natl Acad Sci U S A. 1980 Apr;77(4):2069-73 PMID: 6154943
  3. The primary structure of actin from rabbit skeletal muscle. Three cyanogen bromide peptides that are insoluble at neutral pH.
    J Biol Chem. 1975 Aug 10;250(15):5906-14 PMID: 238990
  4. The amino acid sequence of Physarum actin.
    Nature. 1978 Dec 14;276(5689):720-1 PMID: 732876
  5. Immunization, isolation of immunoglobulins, estimation of antibody titre.
    Scand J Immunol Suppl. 1973;1:161-4 PMID: 4204108
  6. Optical rotatory dispersion of G-actin.
    Biochemistry. 1962 Nov;1:987-96 PMID: 13937183
  7. ATP binding to a protease-resistant core of actin.
    Proc Natl Acad Sci U S A. 1976 Aug;73(8):2742-6 PMID: 134374
  8. Deoxyribonuclease I binding masks the major tryptic cleavage sites on actin.
    Life Sci. 1980 Apr 21;26(16):1323-7 PMID: 6247592
  9. Myogenesis in primary cell cultures from Drosophila melanogaster: protein synthesis and actin heterogeneity during development.
    Cell. 1978 Apr;13(4):589-98 PMID: 418880
  10. Anti-actin specificity of human smooth muscle antibodies in chronic active hepatitis.
    Clin Exp Immunol. 1976 May;24(2):266-72 PMID: 945140
  11. Depolymerization of F-actin by deoxyribonuclease I.
    Cell. 1976 Apr;7(4):531-42 PMID: 133761
  12. The reactivity and function of thiol groups in trout actin.
    Biochem J. 1972 Jan;126(1):21-5 PMID: 4672661
  13. Inducation of antibody against actin from myxomycete plasmodium and its properties.
    Biochemistry. 1975 Jul;14(13):3024-9 PMID: 807241
  14. The dependence of immunological cross-reactivity upon sequence resemblance among lysozymes. I. Micro-complement fixation studies.
    J Biol Chem. 1971 Oct 10;246(19):5978-89 PMID: 4107307
  15. Interaction of immunoglobulin with actin.
    Mol Immunol. 1979 Nov;16(11):881-8 PMID: 583336
  16. Amino acid substitution and the antigenicity of globular proteins.
    Adv Immunol. 1975;20:71-123 PMID: 47221
  17. A competitive radioimmunoassay on a magnetic phase for actin detection.
    FEBS Lett. 1980 Feb 11;110(2):327-9 PMID: 6892803
  18. Selective carbethoxylation of the histidine residues of actin by diethylpyrocarbonate.
    Eur J Biochem. 1974 May 2;44(1):7-12 PMID: 4277645
  19. The measurement of actin concentration in solution: a comparison of methods.
    Anal Biochem. 1974 Nov;62(1):66-74 PMID: 4473917
  20. Three-dimensional structure of the complex of skeletal muscle actin and bovine pancreatic DNAse I at 6-A resolution.
    Proc Natl Acad Sci U S A. 1981 Jul;78(7):4319-23 PMID: 6270671
  21. Anti-actin antibodies of human and rabbit origin.
    Ann Immunol (Paris). 1978 Feb-Mar;129(2-3):245-54 PMID: 98096
  22. Actin from the nematode, Caenorhabditis elegans, is a single electrofocusing species.
    Biochim Biophys Acta. 1977 Aug 23;493(2):304-9 PMID: 889873
  23. Fragmentation of actin by thrombin-like snake venom proteases.
    Biochim Biophys Acta. 1979 Mar 27;577(1):34-43 PMID: 427215
  24. An efficient method to produce specific anti-actin.
    Histochemistry. 1978 Apr 4;55(3):177-84 PMID: 417050
  25. Biochemical and structural characterization of actin from Dictyostelium discoideum.
    J Biol Chem. 1978 Dec 25;253(24):9088-96 PMID: 152763
  26. Vegetative Dictyostelium cells containing 17 actin genes express a single major actin.
    Nature. 1980 Apr 3;284(5755):475-7 PMID: 6892652
  27. Terbium replacement of calcium in carp muscle calcium-binding parvalbumin: an x-ray crystallographic study.
    J Mol Biol. 1975 Jan 15;91(2):229-32 PMID: 1237627
  28. An antiactin antibody that distinguishes between cytoplasmic and skeletal muscle actins.
    J Cell Biol. 1980 Sep;86(3):891-7 PMID: 6893333
  29. Induction of specific anti-actin antibodies and their measurement by ELISA technique.
    J Immunol Methods. 1979;29(2):145-53 PMID: 479613
  30. Change of reactivity of lysine residues upon actin polymerization.
    Biochemistry. 1981 Sep 29;20(20):5914-9 PMID: 6794619
  31. Immunological comparison of muscle actins from mammal, bird and fish: a quantitative approach.
    FEBS Lett. 1979 Jun 1;102(1):69-74 PMID: 88374
  32. The selective blocking of the polymerization reaction of striated muscle actin leading to a derivative suitable for crystallization. Modification of Tyr-53 by 5-diazonium-(1H)tetrazole.
    Eur J Biochem. 1976 Apr 15;64(1):215-8 PMID: 1278154
  33. Proton nuclear magnetic resonance and electron paramagnetic resonance studies on skeletal muscle actin indicate that the metal and nucleotide binding sites are separate.
    Biochemistry. 1980 Dec 9;19(25):5912-6 PMID: 6257295
  34. Localization of actin in Dictyostelium amebas by immunofluorescence.
    J Cell Biol. 1978 Jun;77(3):714-21 PMID: 355262
  35. Effect of tightly bound divalent cation on the equilibria between G-actin-bound and free ATP.
    Eur J Biochem. 1973 Sep 3;37(3):434-40 PMID: 4777250
  36. Actin antibody: the specific visualization of actin filaments in non-muscle cells.
    Proc Natl Acad Sci U S A. 1974 Jun;71(6):2268-72 PMID: 4210210
  37. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide.
    J Mol Biol. 1978 Dec 25;126(4):783-802 PMID: 745245
  38. Tb3+ as a luminescent probe of actin structure: effects of polymerization, KI, and the binding of deoxyribonuclease I.
    Arch Biochem Biophys. 1982 Jun;216(1):81-7 PMID: 6285836
  39. Determination of anti-actin antibodies by a solid-phase immunoenzymatic assay and by indirect immunofluorescence technique.
    Clin Immunol Immunopathol. 1978 Nov;11(3):328-38 PMID: 361319
  40. Crystalline actin tubes. I. Is the conformation of the lanthanide-induced actin tube monomer more like F-actin than G-actin?
    Biochim Biophys Acta. 1980 Jul 24;624(1):163-73 PMID: 6447516
  41. Immunogenicity of proteins and their evolution rate.
    Mol Immunol. 1979 Feb;16(2):123-7 PMID: 221419
  42. Biochemistry of actomyosin-dependent cell motility (a review).
    Proc Natl Acad Sci U S A. 1978 Feb;75(2):588-99 PMID: 147464
  43. 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):4866-71 PMID: 4254541
  44. Actins from mammals, bird, fish and slime mold characterized by isoelectric focusing in polyacrylamide gels.
    Eur J Biochem. 1978 Aug 15;89(1):105-12 PMID: 699900
  45. The actin genes of Drosophila: protein coding regions are highly conserved but intron positions are not.
    Cell. 1981 Apr;24(1):107-16 PMID: 6263481
  46. Differential recognition of monomeric and polymeric forms of actin by anti-actin antibodies.
    Mol Immunol. 1980 Oct;17(10):1219-30 PMID: 6894015
  47. Identification and characterization of multiple forms of actin.
    Cell. 1976 Dec;9(4 PT 2):793-805 PMID: 1017015
  48. Reversible modification of arginine residues. Application to sequence studies by restriction of tryptic hydrolysis to lysine residues.
    J Biol Chem. 1975 Jan 25;250(2):557-64 PMID: 234432
  49. Involvement of an arginine residue of actin in tropomyosin binding.
    Biochem Biophys Res Commun. 1978 Jun 14;82(3):1013-8 PMID: 151536
  50. Co-operativity and calcium/magnesium binding to troponin C and muscle calcium binding parvalbumin: an hypothesis.
    J Theor Biol. 1980 Jun 7;84(3):401-44 PMID: 7431934
  51. Protein-protein interactions of proteolytic fragments of actin.
    Biochim Biophys Acta. 1979 May 23;578(1):253-7 PMID: 156561
  52. The sulfhydryl groups of actin.
    Arch Biochem Biophys. 1968 Jan;123(1):29-40 PMID: 5636367
  53. Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. II. Species-specific and species-non-specific antibodies to smooth and striated chicken muscle actin.
    Histochemistry. 1977 Feb 1;50(4):271-9 PMID: 319085
  54. Conformational integrity of myoglobin after immunization with Freund's adjuvant.
    J Immunol. 1977 Jan;118(1):226-8 PMID: 63516
  55. Myosin in polymorphonuclear leukocytes.
    J Biol Chem. 1973 Dec 10;248(23):8288-94 PMID: 4270951
  56. Interaction between Acanthamoeba actin and rabbit skeletal muscle tropomyosin.
    J Biol Chem. 1977 May 25;252(10):3374-8 PMID: 140871
  57. Detection of antibodies specific to sodium dodecyl sulfate-treated proteins.
    J Immunol Methods. 1979;28(1-2):143-8 PMID: 381529
Article Info
Journal
Journal of muscle research and cell motility
Abbr.
J Muscle Res Cell Motil
ISSN
0142-4319
DOI
10.1007/BF00711947
Published
1983-08-00
Pages
405-27
Language
English
Region
Netherlands
NLM ID
8006298
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00711947 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