Soft metal ions, Cd(II) and Hg(II), induce triple-stranded α-helical assembly and folding of a de novo designed peptide in their trigonal geometries

Xiangqun Li, Kazuo Suzuki, Kenji Kanaori, Kunihiko Tajima, Ayumi Kashiwada, Hidekazu Hiroaki, Daisuke Kohda, Toshiki Tanaka

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

We previously reported the de novo design of an amphiphilic peptide (YGG(IEKKIEA) 4 ) that forms a native-like, parallel triple-stranded coiled coil. Starting from this peptide, we sought to regulate the assembly of the peptide by a metal ion. The replacement of the Ile18 and Ile22 residues with Ala and Cys residues, respectively, in the hydrophobic positions disrupted of the triple-stranded α-helix structure. The addition of Cd(II), however, resulted in the reconstitution of the triple-stranded α-helix bundle, as revealed by circular dichroism (CD) spectroscopy and sedimentation equilibrium analysis. By titration with metal ions and monitoring the change in the intensity of the CD spectra at 222 nm, the dissociation constant K(d) was determined to be 1.5 ± 0.8 μM for Cd(II). The triple-stranded complex formed by the 113 Cd(II) ion showed a single 113 Cd NMR resonance at 572 ppm whose chemical shift was not affected by the presence of Cl - ions. The 113 Cd NMR resonance was connected with the βH protons of the cysteine residue by 1 H- 113 Cd heteronuclear multiple quantum correlation spectroscopy. These NMR results indicate that the three cysteine residues are coordinated to the cadmium ion in a trigonal-planar complex. Hg(II) also induced the assembly of the peptide into a triple-stranded α-helical bundle below the Hg(II)/peptide ratio of 1/3. With excess Hg(II), however, the α- helicity of the peptide was decreased, with the change of the Hg(II) coordination state from three to two. Combining this construct with other functional domains should facilitate the production of artificial proteins with functions controlled by metal ions.

Original languageEnglish
Pages (from-to)1327-1333
Number of pages7
JournalProtein Science
Volume9
Issue number7
DOIs
Publication statusPublished - Jan 1 2000
Externally publishedYes

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Metal ions
Metals
Ions
Peptides
Geometry
Nuclear magnetic resonance
Circular Dichroism
Cysteine
Spectrum Analysis
Circular dichroism spectroscopy
Chemical shift
Dichroism
Cadmium
Titration
Sedimentation
Protons
Spectroscopy
Monitoring
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Soft metal ions, Cd(II) and Hg(II), induce triple-stranded α-helical assembly and folding of a de novo designed peptide in their trigonal geometries. / Li, Xiangqun; Suzuki, Kazuo; Kanaori, Kenji; Tajima, Kunihiko; Kashiwada, Ayumi; Hiroaki, Hidekazu; Kohda, Daisuke; Tanaka, Toshiki.

In: Protein Science, Vol. 9, No. 7, 01.01.2000, p. 1327-1333.

Research output: Contribution to journalArticle

Li, Xiangqun ; Suzuki, Kazuo ; Kanaori, Kenji ; Tajima, Kunihiko ; Kashiwada, Ayumi ; Hiroaki, Hidekazu ; Kohda, Daisuke ; Tanaka, Toshiki. / Soft metal ions, Cd(II) and Hg(II), induce triple-stranded α-helical assembly and folding of a de novo designed peptide in their trigonal geometries. In: Protein Science. 2000 ; Vol. 9, No. 7. pp. 1327-1333.
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abstract = "We previously reported the de novo design of an amphiphilic peptide (YGG(IEKKIEA) 4 ) that forms a native-like, parallel triple-stranded coiled coil. Starting from this peptide, we sought to regulate the assembly of the peptide by a metal ion. The replacement of the Ile18 and Ile22 residues with Ala and Cys residues, respectively, in the hydrophobic positions disrupted of the triple-stranded α-helix structure. The addition of Cd(II), however, resulted in the reconstitution of the triple-stranded α-helix bundle, as revealed by circular dichroism (CD) spectroscopy and sedimentation equilibrium analysis. By titration with metal ions and monitoring the change in the intensity of the CD spectra at 222 nm, the dissociation constant K(d) was determined to be 1.5 ± 0.8 μM for Cd(II). The triple-stranded complex formed by the 113 Cd(II) ion showed a single 113 Cd NMR resonance at 572 ppm whose chemical shift was not affected by the presence of Cl - ions. The 113 Cd NMR resonance was connected with the βH protons of the cysteine residue by 1 H- 113 Cd heteronuclear multiple quantum correlation spectroscopy. These NMR results indicate that the three cysteine residues are coordinated to the cadmium ion in a trigonal-planar complex. Hg(II) also induced the assembly of the peptide into a triple-stranded α-helical bundle below the Hg(II)/peptide ratio of 1/3. With excess Hg(II), however, the α- helicity of the peptide was decreased, with the change of the Hg(II) coordination state from three to two. Combining this construct with other functional domains should facilitate the production of artificial proteins with functions controlled by metal ions.",
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AU - Suzuki, Kazuo

AU - Kanaori, Kenji

AU - Tajima, Kunihiko

AU - Kashiwada, Ayumi

AU - Hiroaki, Hidekazu

AU - Kohda, Daisuke

AU - Tanaka, Toshiki

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