Identification of two sites in gelsolin with different sensitivities to adenine nucleotides

L. E. Laham, M. Way, H. L. Yin, P. A. Janmey

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The affinity of monomeric actin for several actin-binding proteins, including gelsolin, depends on adenine nucleotides. Gelsolin binds faster and with higher affinity to ADP-actin than to ATP-actin. Here, we show that the C-terminal actin-binding domain of gelsolin, which is required for filament nucleating activity but not for filament severing activity, contains the site that distinguishes between ATP-actin and ADP-actin monomers. In contrast, actin binding to the N-terminal half of gelsolin depends on solution ATP concentrations, but not on the nucleotide (ATP or ADP) tightly bound in the cleft of the actin monomer. Binding is stronger in the absence of free nucleotide or in the presence of 0.5 mM ADP than in solutions containing 0.5 mM ATP. Complexes formed using different nucleotide concentrations differ in their filament-severing activities as well as in their abilities to increase the fluorescence of 4-chloro-7-nitrobenzeno-2-oxa-1,3-diazole-labeled actin monomers. These results suggest that, at physiologic concentrations of nucleotides, both free and actin-bound ATP may affect the binding of actin to its accessory proteins and that gelsolin, actin, or the gelsolin-actin complex, contains a low-affinity nucleotide-binding site.

Original languageEnglish (US)
Pages (from-to)1-7
Number of pages7
JournalEuropean Journal of Biochemistry
Volume234
Issue number1
StatePublished - 1995

Fingerprint

Gelsolin
Adenine Nucleotides
Actins
Nucleotides
Adenosine Triphosphate
Monomers
Adenosine Diphosphate
Microfilament Proteins
Accessories
Fluorescence
Binding Sites

Keywords

  • actin
  • ATP
  • gelsolin
  • nucleotides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of two sites in gelsolin with different sensitivities to adenine nucleotides. / Laham, L. E.; Way, M.; Yin, H. L.; Janmey, P. A.

In: European Journal of Biochemistry, Vol. 234, No. 1, 1995, p. 1-7.

Research output: Contribution to journalArticle

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N2 - The affinity of monomeric actin for several actin-binding proteins, including gelsolin, depends on adenine nucleotides. Gelsolin binds faster and with higher affinity to ADP-actin than to ATP-actin. Here, we show that the C-terminal actin-binding domain of gelsolin, which is required for filament nucleating activity but not for filament severing activity, contains the site that distinguishes between ATP-actin and ADP-actin monomers. In contrast, actin binding to the N-terminal half of gelsolin depends on solution ATP concentrations, but not on the nucleotide (ATP or ADP) tightly bound in the cleft of the actin monomer. Binding is stronger in the absence of free nucleotide or in the presence of 0.5 mM ADP than in solutions containing 0.5 mM ATP. Complexes formed using different nucleotide concentrations differ in their filament-severing activities as well as in their abilities to increase the fluorescence of 4-chloro-7-nitrobenzeno-2-oxa-1,3-diazole-labeled actin monomers. These results suggest that, at physiologic concentrations of nucleotides, both free and actin-bound ATP may affect the binding of actin to its accessory proteins and that gelsolin, actin, or the gelsolin-actin complex, contains a low-affinity nucleotide-binding site.

AB - The affinity of monomeric actin for several actin-binding proteins, including gelsolin, depends on adenine nucleotides. Gelsolin binds faster and with higher affinity to ADP-actin than to ATP-actin. Here, we show that the C-terminal actin-binding domain of gelsolin, which is required for filament nucleating activity but not for filament severing activity, contains the site that distinguishes between ATP-actin and ADP-actin monomers. In contrast, actin binding to the N-terminal half of gelsolin depends on solution ATP concentrations, but not on the nucleotide (ATP or ADP) tightly bound in the cleft of the actin monomer. Binding is stronger in the absence of free nucleotide or in the presence of 0.5 mM ADP than in solutions containing 0.5 mM ATP. Complexes formed using different nucleotide concentrations differ in their filament-severing activities as well as in their abilities to increase the fluorescence of 4-chloro-7-nitrobenzeno-2-oxa-1,3-diazole-labeled actin monomers. These results suggest that, at physiologic concentrations of nucleotides, both free and actin-bound ATP may affect the binding of actin to its accessory proteins and that gelsolin, actin, or the gelsolin-actin complex, contains a low-affinity nucleotide-binding site.

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