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PMID: 7543902 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The specific NH2-terminal sequence Ac-EEED of alpha-smooth muscle actin plays a role in polymerization in vitro and in vivo.

The Journal of cell biology ·Vol. 130 ·No. 4 ·1995-08-00 ·Pages 887-95

Chaponnier C, Goethals M, Janmey PA, Gabbiani F, Gabbiani G, Vandekerckhove J

Abstract

The blocking effect of the NH2-terminal decapeptide of alpha-smooth muscle (SM) actin AcEEED-STALVC on the binding of the specific monoclonal antibody anti-alpha SM-1 (Skalli, O., P. Ropraz, A. Trzeviak, G. Benzonana, D. Gillessen, and G. Gabbiani. 1986. J. Cell Biol. 103:2787-2796) was compared with that of synthetic peptides modified by changing the acetyl group or by substituting an amino acid in positions 1 to 5. Using immunofluorescence and immunoblotting techniques, anti-alpha SM-1 binding was abolished by the native peptide and by peptides with a substitution in position 5, indicating that AcEEED is the epitope for anti-alpha SM-1. Incubation of anti-alpha SM-1 (or of its Fab fragment) with arterial SM actin increased polymerization in physiological salt conditions; the antibody binding did not hinder the incorporation of the actin antibody complex into the filaments. This action was not exerted on skeletal muscle actin. After microinjection of the alpha-SM actin NH2-terminal decapeptide or of the epitopic peptide into cultured aortic smooth muscle cells, double immunofluorescence for alpha-SM actin and total actin showed a selective disappearance of alpha-SM actin staining, detectable at approximately 30 min. When a control peptide (e.g. alpha-skeletal [SK] actin NH2-terminal peptide) was microinjected, this was not seen. This effect is compatible with the possibility that the epitopic peptide traps a protein involved in alpha-SM actin polymerization during the dynamic filament turnover in stress fibers. Whatever the mechanism, this is the first evidence that the NH2 terminus of an actin isoform plays a role in the regulation of polymerization in vitro and in vivo.

MeSH Terms
Actins/immunology,metabolism,ultrastructure Amino Acid Sequence Animals Antibody Specificity Aorta/cytology Binding, Competitive Centrifugation Epitopes Immunoblotting Microinjections Microscopy, Electron Microscopy, Fluorescence Molecular Sequence Data Muscle, Smooth, Vascular/metabolism Oligopeptides/immunology,metabolism Peptide Fragments/immunology Polymers/metabolism Protein Binding Rats Rats, Wistar
Chemicals
Actins Epitopes Oligopeptides Peptide Fragments Polymers
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Chaponnier C
Department of Pathology, University of Geneva, Switzerland.
Goethals M
Janmey P A
Gabbiani F
Gabbiani G
Vandekerckhove J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.130.4.887
Published
1995-08-00
Pages
887-95
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2199961
Subset
IM
Grants
NIAMS NIH HHS · AR38910 · United States
External Links
PubMed source DOI: 10.1083/jcb.130.4.887 PMC Full Text View journal details
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