4G1A

Metal-binding properties of a self-assembled coiled coil: formation of a polynuclear Cd-thiolated cluster

Summary for 4G1A

• Entry DOI: 10.2210/pdb4g1a/pdb
• Descriptor: AQ-C16C19 peptide, CADMIUM ION (3 entities in total)
• Functional Keywords: helical bundles, metallopeptide complexes, polynuclear metal-binding, cd(ii), self-assembly, metal binding protein
• Biological source: synthetic construct
• Total number of polymer chains: 3
• Total formula weight: 10718.54

Authors

Ni, S.,Kennedy, M.A.,Ogawa, M.Y. (deposition date: 2012-07-10, release date: 2012-12-05, Last modification date: 2024-02-28)

Primary citation

Zaytsev, D.V.,Morozov, V.A.,Fan, J.,Zhu, X.,Mukherjee, M.,Ni, S.,Kennedy, M.A.,Ogawa, M.Y.
Metal-binding properties and structural characterization of a self-assembled coiled coil: Formation of a polynuclear Cd-thiolate cluster.
J.Inorg.Biochem., 119C:1-9, 2012

PubMed Abstract: This paper describes the design, characterization, and metal-binding properties of a 32-residue polypeptide called AQ-C16C19. The sequence of this peptide is composed of four repeats of the seven residue sequence Ile-Ala-Ala-Leu-Glu-Gln-Lys but with a Cys-X-X-Cys metal-binding motif substituted at positions 16-19. Size exclusion chromatography with multiangle light scattering detection (SEC-MALS) and circular dichroism (CD) spectroscopy studies showed that the apo peptide exhibits a pH-dependent oligomerization state in which a three-stranded α-helical coiled coil is dominant between pH5.4 and 8.5. The Cd(2+)-binding properties of the AQ-C16C19 peptide were studied by ultraviolet-visible spectroscopy (UV-vis), electrospray ionization mass spectrometry (ESI MS), and (113)Cd NMR techniques. The holoprotein was found to contain a polynuclear cadmium-thiolate center formed within the hydrophobic core of the triple-stranded α-helical coiled-coil structure. The X-ray crystal structure of the Cd-loaded peptide, resolved at 1.85Å resolution, revealed an adamantane-like configuration of the polynuclear metal center consisting of four cadmium ions, six thiolate sulfur ligands from cysteine residues and four oxygen-donor ligands. Three of these are from glutamic acid residues and one is from an exogenous water molecule. Thus, each cadmium ion is coordinated in a distorted tetrahedral S(3)O geometry. The metal cluster was found to form cooperatively at pH5.4 but in a stepwise fashion at pH>7. The results demonstrate that synthetic coiled-coils can be designed to incorporate multinuclear metal clusters, a proof-of-concept for their potential use in developing synthetic metalloenzymes and multi-electron redox agents.

PubMed: 23160144
DOI: 10.1016/j.jinorgbio.2012.10.010

Experimental method

X-RAY DIFFRACTION (1.85 Å)