Evidence that a 27-residue sequence is the actin-binding site of ABP-120

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Abstract

Proteolysis experiments of ABP-120 from Dictyostelium discoideum have previously demonstrated that removal of residues 89-115 from a tryptic peptide which retains actin binding activity, abolishes actin binding (Bresnick, A. R., Warren, V., and Condeelis, J. (1990) J. Biol. Chem. 265, 9236-9240). Antibodies made against a synthetic peptide of this 27-amino acid sequence (27-mer) specifically immunoprecipitate native ABP-120 from Dictyostelium high speed supernatants, demonstrating that the 27-mer sequence is on the surface of the molecule as expected for an active rite. ABP-120 is inhibited in its binding to F-actin by Fab′ fragments of the anti-27-mer IgG. Half-maximal inhibition occurs at an approximate molar ratio of 7 Fab′ fragments/ABP-120 monomer. Viscoelastic measurements indicate that ABP-120 forms fewer crosslinks with F-actin in the presence of the 27-mer synthetic peptide than in its absence. In F-actin cosedimentation assays, the binding of ABP-120 to actin is inhibited by the 27-mer synthetic peptide. Furthermore, the 27-mer synthetic peptide cosediments with F-actin, whereas a control hydrophobic peptide and a synthetic peptide of residues 69-88 of ABP-120 do not cosediment with F-actin. These observations suggest a direct involvement of the 27-mer sequence in the actin binding activity of ABP-120.

Original languageEnglish (US)
Pages (from-to)12989-12993
Number of pages5
JournalJournal of Biological Chemistry
Volume266
Issue number20
StatePublished - 1991

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Actins
Binding Sites
Peptides
Immunoglobulin Fab Fragments
Dictyostelium
Proteolysis
Amino Acid Sequence
Assays
Immunoglobulin G
Monomers
Amino Acids
Molecules
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Evidence that a 27-residue sequence is the actin-binding site of ABP-120. / Bresnick, Anne R.; Janmey, Paul A.; Condeelis, John S.

In: Journal of Biological Chemistry, Vol. 266, No. 20, 1991, p. 12989-12993.

Research output: Contribution to journalArticle

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