Time-dependent, protein-directed growth of gold nanoparticles within a single crystal of lysozyme

Journal name:
Nature Nanotechnology
Volume:
6,
Pages:
93–97
Year published:
DOI:
doi:10.1038/nnano.2010.280
Received
Accepted
Published online

Abstract

Gold nanoparticles are useful in biomedical applications due to their distinct optical properties and high chemical stability1, 2, 3, 4, 5. Reports of the biogenic formation of gold colloids from gold complexes has also led to an increased level of interest in the biomineralization of gold6, 7, 8, 9, 10, 11, 12, 13. However, the mechanism responsible for biomolecule-directed gold nanoparticle formation remains unclear due to the lack of structural information about biological systems and the fast kinetics of biomimetic chemical systems in solution. Here we show that intact single crystals of lysozyme can be used to study the time-dependent, protein-directed growth of gold nanoparticles. The protein crystals slow down the growth of the gold nanoparticles, allowing detailed kinetic studies to be carried out, and permit a three-dimensional structural characterization that would be difficult to achieve in solution. Furthermore, we show that additional chemical species can be used to fine-tune the growth rate of the gold nanoparticles.

At a glance

Figures

  1. Time-dependent, protein-directed growth of gold nanoparticles in single crystals.
    Figure 1: Time-dependent, protein-directed growth of gold nanoparticles in single crystals.

    a, Optical images of single crystals of lysozyme grown in the presence of Au(I) at different days of growth. bd, Corresponding TEM images at low magnification (b) and high magnification (c), and plots of the size distribution histograms (d) of the gold nanoparticles within the lysozyme crystals (s.d., standard deviation). e, X-ray crystal structures of the lysozymes from single crystals in a at the first, second, third and 90th days of growth. (Au(I), ochre; Au(III), green; carbon, cyan; nitrogen, blue; oxygen, red; chloride, magenta.) f, Time-dependent size evolution of gold nanoparticles based on their TEM images. g, Time-dependent occupancy evolution of different gold ions bound to the three-dimensional structure of lysozymes. The numbering scheme of gold ions is the same as in e.

  2. HAADF-STEM images of the three-dimensional distribution of gold nanoparticles within lysozyme single crystals.
    Figure 2: HAADF-STEM images of the three-dimensional distribution of gold nanoparticles within lysozyme single crystals.

    a, HAADF-STEM image of gold nanoparticles within lysozyme single crystals at 0° tilt. b, Corresponding image of the three-dimensional tomographic reconstruction. c, As in b, but without lysozyme. In a, white dots indicate gold nanoparticles incorporated within the lysozyme matrix, which is slightly darker. In b and c, gold nanoparticles are in yellow and lysozyme crystals are in blue.

  3. Schematic of gold nanoparticle growth within a lysozyme single crystal.
    Figure 3: Schematic of gold nanoparticle growth within a lysozyme single crystal.

    a, Lysozyme (blue) and Au(I) (ochre) are mixed to allow the protein crystals to grow. b, Au(I) is initially bound specifically to the ε-N of His15 of the lysozyme, and then dissociates from His15 and disproportionates into Au(0) (yellow) and Au(III) (green). Meanwhile, Au(I) in the mother liquor diffuses continuously into the lysozyme crystals and then disproportionates into Au(0) and Au(III). Au(0) then grows into small gold clusters (the gold clusters are too small to render the crystal red at this early stage) and the Au(III) translocates and rebinds the lysozyme. c, As the process proceeds, more Au(I) disproportionates into Au(0), making the small gold clusters grow larger, into nanoparticles, rendering the crystal red due to their characteristic SPR absorption. At the same time, more Au(III) are generated from the disproportionation and translocate and rebind the lysozyme. d, Finally, Au(I) disappears and a large number of ~20 nm gold nanoparticles form within the crystal, while eight Au(III) rebind to the lysozyme. The enlarged pictures in bd show experimentally determined crystallographic packing of lysozyme proteins with gold ions inside. For clarity of the scheme, the lysozyme proteins are not drawn to scale in the three-dimensional protein crystals.

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Affiliations

  1. Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA

    • Hui Wei,
    • Limin Yang,
    • Li Huey Tan,
    • Hang Xing &
    • Yi Lu
  2. Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA

    • Zidong Wang,
    • Jiong Zhang,
    • Stephen House,
    • Ian M. Robertson,
    • Jian-Min Zuo &
    • Yi Lu
  3. George L. Clark X-Ray Facility and 3M Materials Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA

    • Yi-Gui Gao
  4. College of Bioengineering, Chongqing University, Sha Ping Ba Street 174, Chongqing, 400044, China

    • Limin Yang &
    • Changjun Hou
  5. Department of Biology, Brookhaven National Laboratory, Upton, New York 11973, USA

    • Howard Robinson

Contributions

H.W., Z.W. and Y.L. designed the research. H.W., Z.W., J.Z., Y.-G.G., L.Y. and H.R. performed the research. H.W., Z.W., J.Z., S.H., Y.-G.G., L.Y., H.R., L.H.T., H.X., C.H., I.M.R., J.-M.Z. and Y.L. analysed the data. All authors co-wrote the paper.

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