Targeting the Surface of the Protein 14‐3‐3 by Ultrasmall (1.5 nm) Gold Nanoparticles Carrying the Specific Peptide CRaf

Ruks, Tatjana; Loza, Kateryna; Heggen, Marc; Ottmann, C.; Bayer, Peter; Beuck, Christine; Epple, Matthias

doi:10.1002/cbic.202000761

Cited by 12 publications

(11 citation statements)

References 57 publications

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“…a molecular footprint of 0.074 nm 2 per cysteine in cysteine-bearing precision macromolecules 70 and of 0.41 nm 2 for the peptide CRaf with 36 amino acids, bound via a cysteine. 71 For all subsequent calculations and considerations, we assumed that the peptide concentration c(peptide) was 150 times the gold nanoparticle concentration c(AuNP).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Peptide-Conjugated Ultrasmall Gold Nanoparticles (2 nm) for Selective Protein Targeting

Ruks

Loza

Heggen

et al. 2021

ACS Appl. Bio Mater.

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Ultrasmall gold nanoparticles with a metallic core diameter of 2 nm were surface-conjugated with peptides that selectively target epitopes on the surface of the WW domain of the model protein hPin1 (hPin1-WW). The binding to the gold surface was accomplished via the thiol group of a terminal cysteine. The particles were analyzed by NMR spectroscopy, highresolution transmission electron microscopy, and differential centrifugal sedimentation. The surface loading was determined by conjugating a FAMlabeled peptide, followed by UV−vis spectroscopy, and by quantitative 1 H NMR spectroscopy, showing about 150 peptide molecules conjugated to each nanoparticle. The interaction between the peptide-decorated nanoparticles with hPin1-WW was probed by 1 H− 15 N-HSQC NMR titration, fluorescence polarization spectroscopy (FP), and isothermal titration calorimetry (ITC). The particles showed a similar binding (K D = 10−20 μM) compared to the dissolved peptides (K D = 10−30 μM). Small-angle Xray scattering (SAXS) showed that the particles were well dispersed and did not agglomerate after the addition of hPin1-WW (no cross-linking by the protein). Each nanoparticle was able to bind about 20 hPin1-WW protein molecules. An unspecific interaction with hPin1 was excluded by the attachment of a nonbinding peptide to the nanoparticle surface. The uptake by cells was studied by confocal laser scanning microscopy. The peptide-functionalized nanoparticles penetrated the cell membrane and were located in the cytosol. In contrast, the dissolved peptide did not cross the cell membrane. Peptide-functionalized nanoparticles are promising agents to target proteins inside cells.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Peptide-Conjugated Ultrasmall Gold Nanoparticles (2 nm) for Selective Protein Targeting

Ruks

Loza

Heggen

et al. 2021

ACS Appl. Bio Mater.

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“…Nevertheless, peptides can be easily attached via cysteine to serve as capping agent and targeting molecule at the same time. 50,51 For more sensitive ligands and for a better control over stoichiometry, it is preferable to attach the ligand after the particle has been prepared and stabilized. Ligand-exchange reactions 49 and versatile conjugation methods, e.g.…”

Section: Nanoparticles For Targeting and Interaction With Biomolecule...mentioning

confidence: 99%

The Why and How of Ultrasmall Nanoparticles

Epple,

Rotello,

Dawson

2023

Acc. Chem. Res.

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In this Account, we describe our research into ultrasmall nanoparticles, including their unique properties, and outline some of the new opportunities they offer. We will summarize our perspective on the current state of the field and highlight what we see as key questions that remain to be solved. First, there are several nanostructure size-scale regimes, with qualitatively distinct functional biological attributes. Broadly generalized, larger particles (e.g., larger than 300 nm) tend to be more efficiently swept away by the first line of the immune system (for example macrophages). In the "middle-sized" regime (20−300 nm), nanoparticle surfaces and shapes can be recognized by energy-dependent cellular reorganizations, then organized locally in a spatial and temporally coherent way. That energy is gated and made available by specific cellular recognition processes. The relationship between particle surface design, endogenously derived nonspecific biomolecular corona, and architectural features recognized by the cell is complex and only purposefully and very precisely designed nanoparticle architectures are able to navigate to specific targets. At sufficiently small sizes (<10 nm including the ligand shell, associated with a core diameter of a few nm at most) we enter the "quasimolecular regime" in which the endogenous biomolecular environment exchanges so rapidly with the ultrasmall particle surface that larger scale cellular and immune recognition events are often greatly simplified. As an example, ultrasmall particles can penetrate cellular and biological barriers within tissue architectures via passive diffusion, in much the same way as small molecule drugs do. An intriguing question arises: what happens at the interface of cellular recognition and ultrasmall quasi-molecular size regimes? Succinctly put, ultrasmall conjugates can evade defense mechanisms driven by larger scale cellular nanoscale recognition, enabling them to flexibly exploit molecular interaction motifs to interact with specific targets. Numerous advances in control of architecture that take advantage of these phenomena have taken place or are underway. For instance, syntheses can now be sufficiently controlled that it is possible to make nanoparticles of a few hundreds of atoms or metalloid clusters of several tens of atoms that can be characterized by single crystal X-ray structure analysis. While the synthesis of atomically precise clusters in organic solvents presents challenges, water-based syntheses of ultrasmall nanoparticles can be upscaled and lead to well-defined particle populations. The surface of ultrasmall nanoparticles can be covalently modified with a wide variety of ligands to control the interactions of these particles with biosystems, as well as drugs and fluorophores. And, in contrast to larger particles, many advanced molecular analytical and separation tools can be applied to understand their structure. For example, NMR spectroscopy allows us to obtain a detailed image of the particle surface and the attached ligands. The...

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“…Currently, ensemble methods like Western blot, BCA assay and ζ-potential are used as indicators for changes in surface physical features and biomolecule functionalization of materials. [75][76][77][78][79] By using STORM or DNA-PAINT, these characteristics can be resolved on a single molecule level. Later, counting of integer numbers of molecules can be achieved.…”

Section: Informative and Quantitative Outputmentioning

confidence: 99%

Can super-resolution microscopy become a standard characterization technique for materials chemistry?

et al. 2022

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Targeting the Surface of the Protein 14‐3‐3 by Ultrasmall (1.5 nm) Gold Nanoparticles Carrying the Specific Peptide CRaf

Cited by 12 publications

References 57 publications

Peptide-Conjugated Ultrasmall Gold Nanoparticles (2 nm) for Selective Protein Targeting

Peptide-Conjugated Ultrasmall Gold Nanoparticles (2 nm) for Selective Protein Targeting

The Why and How of Ultrasmall Nanoparticles

Can super-resolution microscopy become a standard characterization technique for materials chemistry?

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