Two-dimensional crystals of streptavidin on biotinylated lipid layers and their interactions with biotinylated macromolecules

Darst, Seth A.; Ahlers, Michael; Meller, P.; Kubalek, Elizabeth W.; Blankenburg, R.; Ribi, Hans O.; Ringsdorf, Helmut; Kornberg, Roger D.

doi:10.1016/s0006-3495(91)82232-9

Cited by 328 publications

(359 citation statements)

References 18 publications

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“…Such crystals display two accessible biotin-binding sites per tetramer that can be used to bind biotinylated proteins (26). However, contrary to streptavidin, S-layer proteins form two-dimensional protein crystals by a robust self-assembly process that has been optimized by evolution.…”

Section: Discussionmentioning

confidence: 99%

S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays

Moll¹,

Huber²,

Schlegel³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

210

196

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Biomolecular self-assembly can be used as a powerful tool for nanoscale engineering. In this paper, we describe the development of building blocks for nanobiotechnology, which are based on the fusion of streptavidin to a crystalline bacterial cell surface layer (S-layer) protein with the inherent ability to self-assemble into a monomolecular protein lattice. The fusion proteins and streptavidin were produced independently in Escherichia coli, isolated, and mixed to refold and purify heterotetramers of 1:3 stoichiometry. Self-assembled chimeric S-layers could be formed in suspension, on liposomes, on silicon wafers, and on accessory cell wall polymer containing cell wall fragments. The two-dimensional protein crystals displayed streptavidin in defined repetitive spacing, and they were capable of binding D-biotin and biotinylated proteins. Therefore, the chimeric S-layer can be used as a self-assembling nanopatterned molecular affinity matrix to arrange biotinylated compounds on a surface. In addition, it has application potential as a functional coat of liposomes.

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Section: Discussionmentioning

confidence: 99%

S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays

Moll¹,

Huber²,

Schlegel³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

210

196

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“…The two-dimensional crystals of streptavidin can easily be formed on highly fluidic PLBs containing lipids with unsaturated alkyl chains (such as DOPC, dioleoylphosphatidylcholine) and a biotin-containing lipid (11,20). Their surface is particularly useful for the selective and stable immobilization of homo-oligomeric protein complexes because pinning the complexes at multiple biotinylated sites is possible (109).…”

Section: Substrate Surfacesmentioning

confidence: 99%

High-Speed AFM and Applications to Biomolecular Systems

Ando

Uchihashi

Kodera

2013

Annu. Rev. Biophys.

252

216

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Directly observing individual protein molecules in action at high spatiotemporal resolution has long been a holy grail for biological science. This is because we long have had to infer how proteins function from the static snapshots of their structures and dynamic behavior of optical makers attached to the molecules. This limitation has recently been removed to a large extent by the materialization of high-speed atomic force microscopy (HS-AFM). HS-AFM allows us to directly visualize the structure dynamics and dynamic processes of biological molecules in physiological solutions, at subsecond to sub-100-ms temporal resolution, without disturbing their function. In fact, dynamically acting molecules such as myosin V walking on an actin filament and bacteriorhodopsin in response to light are successfully visualized. In this review, we first describe theoretical considerations for the highest possible imaging rate of this new microscope, and then highlight recent imaging studies. Finally, the current limitation and future challenges to explore are described.

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“…As discussed above, by linking the dyes to the AuNRs surface with the biotin/streptavidin conjugation, they are kept at an average distance of 5 nm from the metal, thus, avoiding the quenching of the plasmonicfluorescent nanoprobes. 30,31 At this distance, the still-appreciable field enhancement produced by the AuNR (see Figure 2 To demonstrate the potential of the AuNRs for NP-STED, we functionalized a glass cover-slip to attach the AuNRs to the surface (see Methods for further details) and compared the resolution improvement relative to confocal microscopy for the AuNR and for 20 nm diameter crimson red beads as a function of the power of the depletion-beam. Assuming that the depletion beam pulse length, , is much smaller than the fluorescence lifetime of the fluorophore, 1 , the resolution achievable with pulsed STED depends on the wavelength, λ, the numerical aperture (NA) of the objective, the peak intensity of the depletion beam, , and the saturation intensity of the fluorophore, , as given by: 16,32 Hence, we can write the STED resolution improvement with respect to confocal microscopy as Γ = √1 + ∅.…”

Section: Green (Blue) Line Is the Absorption (Emission) Spectrum Of Tmentioning

confidence: 99%

Plasmonic Nanoprobes for Stimulated Emission Depletion Nanoscopy

Cortés¹,

Huidobro²,

Sinclair³

et al. 2016

ACS Nano

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Plasmonic nanoparticles influence the absorption and emission processes of nearby emitters due to local enhancements of the illuminating radiation and the photonic density of states. Here, we use the plasmon resonance of metal nanoparticles in order to enhance the stimulated depletion of excited molecules for super-resolved nanoscopy. We demonstrate stimulated emission depletion (STED) nanoscopy with gold nanorods with a long axis of only 26 nm and a width of 8 nm that provide an enhancement of up to 50% of the resolution compared to fluorescent-only probes without plasmonic components irradiated with the same depletion power. The nanoparticle-assisted STED probes reported here represent a ~2x10 3 reduction in probe volume compared to previously used nanoparticles. Finally, we demonstrate their application toward plasmon-assisted STED cellular imaging at low-depletion powers and we also discuss their current limitations. Key-words: STED nanoscopy, nanorods, super-resolution, plasmonic nanoparticles, bio-imagingThe diffraction limit has ceased to be a practical limit to resolution in far-field microscopy, following the demonstration of STED, 1,2,3 RESOLFT 4 and localisation microscopies 5,6,7 and the subsequent development of a plethora of super-resolved nanoscopy techniques. 8 In particular, stimulated emission depletion (STED) nanoscopy, which builds on the advantages of laser scanning confocal microscopy, is a powerful technique for super-resolved imaging in complex biological samples including live organisms. 9,10 STED nanoscopy uses stimulated emission to turn off the spontaneous fluorescence emission of dye molecules, typically overlapping a focused excitation beam with a "doughnut" shaped beam that deexcites emitters to the ground state everywhere except for the area within the centre of the doughnut, thus providing theoretically diffraction-unlimited resolution in the transverse plane by reducing the fullwidth half-maximum (FWHM) of the point spread function. By increasing the power of the depletion beam the emission region can be can be drastically reduced -theoretically allowing for sub-nanometre resolution -with resolutions of less than 10 nm being demonstrated. 11 The scaling of resolution with the square root of the depletion beam power means that relatively high-power lasers are typically used for STED nanoscopy. In practice, however, the use of high-power irradiation can result in problems such as photobleaching of the fluorophores and phototoxicity, and so the achievable resolution is compromised by the need to limit the intensity of the depletion laser radiation. Furthermore, high power lasers can add cost and complexity to STED microscopes and so the requirement for high power depletion beams presents challenges for parallelizing STED measurements 12,13 in terms of the lasers required, thus, limiting the potential for faster super-resolved imaging.To some extent the issue of photobleaching can be addressed with the development of more robust fluorescent labels such as quantum dots 14 as ...

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Two-dimensional crystals of streptavidin on biotinylated lipid layers and their interactions with biotinylated macromolecules

Cited by 328 publications

References 18 publications

S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays

S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays

High-Speed AFM and Applications to Biomolecular Systems

Plasmonic Nanoprobes for Stimulated Emission Depletion Nanoscopy

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