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1.
Opt Express ; 32(6): 8506-8519, 2024 Mar 11.
Article in English | MEDLINE | ID: mdl-38571108

ABSTRACT

In this paper, a 1 × 2 photonic switch is designed based on a silicon-on-insulator (SOI) platform combined with the phase change material (PCM), Sb2S3, assisted by the direct binary search (DBS) algorithm. The designed photonic switch exhibits an impressive operating bandwidth ranging from 1450 to 1650 nm. The device has an insertion loss (IL) from 0.44 dB to 0.70 dB (of less than 0.7 dB) and cross talk (CT) from -26 dB to -20 dB (of less than -20 dB) over an operating bandwidth of 200 nm, especially an IL of 0.52 dB and CT of -24 dB at 1550 nm. Notably, the device is highly compact, with footprints of merely 3 × 4 µm2. Furthermore, we have extended the device's functionality for multifunctional operation in the C-band that can serve as both a 1 × 2 photonic switch and a 3 dB photonic power splitter. In the photonic switch mode, the device demonstrates an IL of 0.7 dB and a CT of -13.5 dB. In addition, when operating as a 3 dB photonic power splitter, the IL is less than 0.5 dB.

2.
Opt Express ; 32(3): 3379-3393, 2024 Jan 29.
Article in English | MEDLINE | ID: mdl-38297560

ABSTRACT

In contrast to conventional emitters fashioned from traditional materials, tunable thermal emitters exhibit a distinct propensity to fulfill the demands of diverse scenarios, thereby engendering an array of prospects within the realms of communications, military applications, and control systems. In this paper, a tunable thermal emitter without continuous external excitation is introduced using Ge2Sb2Te5 (GST) and high-temperature-resistant material Mo. It is automatically optimized by inverse design with genetic algorithm (GA) to switch between different functions according to the object temperature to adapt to diverse scenarios. In "off" mode, the emitter orchestrates a blend of infrared (IR) stealth and thermal management. This is evidenced by average absorptivity values of 0.08 for mid-wave infrared (MIR, 3-5 µm), 0.19 for long-wave infrared (LIR, 8-14 µm), and 0.68 for the non-atmospheric window (NAW, 5-8 µm). Conversely, when confronted with high-temperature entities, the emitter seamlessly transitions to "on" mode, instigating a process of radiative cooling. This transformation is reflected in the augmented emissivity of the dual-band atmospheric window including MIR and LIR, attaining peak values of 0.96 and 0.97. This transition yields a cooling potential, quantified at 64 W/m2 at the ambient temperature of 25°C. In addition, our design employs a layered structure, which avoids complex patterned resonators and facilitates large-area fabrication. The emitter in this paper evinces robust insensitivity to polarization variations and the angle of incidence. We believe that this work will contribute to the development in the fields of dynamic tunability for IR stealth, dynamic radiative cooling systems, and thermal imaging.

3.
Opt Express ; 31(22): 35653-35669, 2023 Oct 23.
Article in English | MEDLINE | ID: mdl-38017732

ABSTRACT

In recent years, structural color has developed rapidly due to its distinct advantages, such as low loss, high spatial resolution and environmental friendliness. Various inverse design methods have been extensively investigated to efficiently design optical structures. However, the optimization method for the inverse design of structural color remains a formidable challenge. Traditional optimization approaches, such as genetic algorithms require time-consuming repetitions of structural simulations. Deep learning-assisted design necessitates prior simulations and large amounts of data, making it less efficient for systems with a small number of features. This study proposes a tensor completion algorithm capable of swiftly and accurately predicting missing datasets based on partially obtained datasets to assist in structural color design. Transforming the complex physical problem of structural color design into a spatial structure relationship problem linking geometric parameters and spectral data. The method utilizes tensor multilinear data analysis to effectively capture the complex relationships associated with geometric parameters and spectral data in higher-order data. Numerical and experimental results demonstrate that the algorithm exhibits high reliability in terms of speed and accuracy for diverse structures, datasets of varying sizes, and different materials, significantly enhancing design efficiency. The proposed algorithm offers a viable solution for inverse design problems involving complex physical systems, thereby introducing a novel approach to the design of photonic devices. Additionally, numerical experiments illustrate that the structural color of cruciform resonators with diamond can overcome the high loss issues observed in traditional dielectric materials within the blue wavelength region and enhance the corrosion resistance of the structure. We achieve a wide color gamut and a high-narrow reflection spectrum nearing 1 by this structure, and the theoretical analysis further verifies that diamond holds great promise in the realm of optics.

4.
Opt Express ; 31(20): 33622-33637, 2023 Sep 25.
Article in English | MEDLINE | ID: mdl-37859139

ABSTRACT

Infrared (IR) thermal camouflage and management are deeply desirable in the field of military and astronomy. While IR compatible with laser camouflage technology is extensively studied to counter modern detection systems, most existing strategies for visible light camouflage focus on color matching, which is not suitable for scenarios requiring transparency. In this work, we propose an optically transparent metamaterial with multi-band compatible camouflage capability based on the inverse design. The metamaterial consists of Ag grating, Si3N4 dielectric spacer layer, Ag reflection layer, and Si3N4 anti-reflective layer. An ideal multi-band compatible spectrum is involved in the inverse design algorithm. Calculated results demonstrate high transmittance (T0.38-0.78µm = 0.70) in the visible region, low reflectance (R1.55µm = 0.01) in laser working wavelength, high reflectance (R3-5µm = 0.86 and R8-14µm = 0.92) in the dual-band atmospheric window, and high emissivity (ɛ5-8µm = 0.61) for the non-atmospheric window. The radiative heat flux in the detected band is 31W/m2 and 201W/m2 respectively. Furthermore, the incident and polarized insensitivity of the proposed metamaterial supports applicability for practical situations. This work, emphasizes an effective strategy for conducting optically transparent design with compatible IR-laser camouflage as well as radiative cooling properties by an automated design approach.

5.
Opt Express ; 31(18): 29235-29244, 2023 Aug 28.
Article in English | MEDLINE | ID: mdl-37710728

ABSTRACT

In this work, we use the inverse design method to design three-channel and four-channel dual-mode waveguide crossings with the design regions of 4.32 µm-wide regular hexagon and 6.68 µm-wide regular octagon, respectively. Based on the highly-symmetric structures, the fundamental transverse electric (TE0) and TE1 modes propagate through the waveguide crossings efficiently. Moreover, the devices are practically fabricated and experimentally characterized. The measured insertion losses and crosstalks of the three-channel and dual-mode waveguide crossing for both the TE0 and TE1 modes are less than 1.8 dB and lower than -18.4 dB from 1540 nm to 1560 nm, respectively. The measured insertion losses of the four-channel and dual-mode waveguide crossing for the TE0 and TE1 modes are less than 1.8 dB and 2.5 dB from 1540 nm to 1560 nm, respectively, and the measured crosstalks are lower than -17.0 dB. In principle, our proposed scheme can be extended to waveguide crossing with more channels and modes.

6.
Opt Express ; 31(17): 27393-27406, 2023 Aug 14.
Article in English | MEDLINE | ID: mdl-37710816

ABSTRACT

In this work, we design, fabricate, and characterize a different-mode (waveguide-connected) power splitter ((W)PS) by what we believe to be a novel multi-dimension direct-binary-search algorithm that can significantly balance the device performance, time cost, and fabrication robustness by searching the state-dimension, rotation-dimension, shape-dimension, and size-dimension parameters. The (W)PS can simultaneously generate the fundamental transverse electric (TE0) and TE1 mode with the 1:1 output balance. Compared with the PS, the WPS can greatly shorten the adiabatic taper length between the single-mode waveguide and the grating coupler. The measured results of the different-mode (W)PS indicate that the insertion loss and crosstalk are less than 0.9 (1.3) dB and lower than -17.8 (-14.9) dB from 1540 nm to 1560 nm. In addition, based on the tunable tap couplers, the different-mode (W)PS can be extended to multiple output ports with different modes and different transmittances.

7.
Opt Express ; 31(11): 18555-18566, 2023 May 22.
Article in English | MEDLINE | ID: mdl-37381565

ABSTRACT

Blindly increasing the channels of the mode (de)multiplexer on the single-layer chip can cause the device structure to be too complex to optimize. The three-dimensional (3D) mode division multiplexing (MDM) technology is a potential solution to extend the data capacity of the photonic integrated circuit by assembling the simple devices in the 3D space. In our work, we propose a 16 × 16 3D MDM system with a compact footprint of about 100 µm × 5.0 µm × 3.7 µm. It can realize 256 mode routes by converting the fundamental transverse electric (TE0) modes in arbitrary input waveguides into the expected modes in arbitrary output waveguides. To illustrate its mode-routing principle, the TE0 mode is launched in one of the sixteen input waveguides, and converted into corresponding modes in four output waveguides. The simulated results indicate that the ILs and CTs of the 16 × 16 3D MDM system are less than 3.5 dB and lower than -14.2 dB at 1550 nm, respectively. In principle, the 3D design architecture can be scaled to realize arbitrary network complexity levels.

8.
Nanomaterials (Basel) ; 13(9)2023 Apr 24.
Article in English | MEDLINE | ID: mdl-37176996

ABSTRACT

Color displays have become increasingly attractive, with dielectric optical nanoantennas demonstrating especially promising applications due to the high refractive index of the material, enabling devices to support geometry-dependent Mie resonance in the visible band. Although many structural color designs based on dielectric nanoantennas employ the method of artificial positive adjustment, the design cycle is too lengthy and the approach is non-intelligent. The commonly used phase change material Ge2Sb2Te5 (GST) is characterized by high absorption and a small contrast to the real part of the refractive index in the visible light band, thereby restricting its application in this range. The Sb2S3 phase change material is endowed with a wide band gap of 1.7 to 2 eV, demonstrating two orders of magnitude lower propagation loss compared to GST, when integrated onto a silicon waveguide, and exhibiting a maximum refractive index contrast close to 1 at 614 nm. Thus, Sb2S3 is a more suitable phase change material than GST for tuning visible light. In this paper, genetic algorithms and finite-difference time-domain (FDTD) solutions are combined and introduced as Sb2S3 phase change material to design nanoantennas. Structural color is generated in the reflection mode through the Mie resonance inside the structure, and the properties of Sb2S3 in different phase states are utilized to achieve tunability. Compared to traditional methods, genetic algorithms are superior-optimization algorithms that require low computational effort and a high population performance. Furthermore, Sb2S3 material can be laser-induced to switch the transitions of the crystallized and amorphous states, achieving reversible color. The large chromatic aberration ∆E modulation of 64.8, 28.1, and 44.1 was, respectively, achieved by the Sb2S3 phase transition in this paper. Moreover, based on the sensitivity of the structure to the incident angle, it can also be used in fields such as angle-sensitive detectors.

9.
Opt Express ; 31(5): 8668-8681, 2023 Feb 27.
Article in English | MEDLINE | ID: mdl-36859977

ABSTRACT

Metalens with extended depth of focus (EDOF) can extend the mapping area of the image, which leads to novel applications in imaging and microscopy. Since there are still some disadvantages for existing EDOF metalenses based on forward design, such as asymmetric point spread function (PSF) and non-uniformly distributed focal spot, which impair the quality of images, we propose a double-process genetic algorithm (DPGA) optimization to inversely design the EDOF metalens for addressing these drawbacks. By separately adopting different mutation operators in successive two genetic algorithm (GA) processes, DPGA exhibits significant advantages in searching for the ideal solution in the whole parameter space. Here, the 1D and 2D EDOF metalenses operating at 980 nm are separately designed via this method, and both of them exhibit significant depth of focus (DOF) improvement to that of conventional focusing. Furthermore, a uniformly distributed focal spot can be maintained well, which can guarantee stable imaging quality along the longitudinal direction. The proposed EDOF metalenses have considerable potential applications in biological microscopy and imaging, and the scheme of DPGA can be promoted to the inverse design of other nanophotonics devices.

10.
Biophys J ; 121(16): 3049-3060, 2022 08 16.
Article in English | MEDLINE | ID: mdl-35841142

ABSTRACT

Intrinsically disordered proteins (IDPs) play critical roles in regulatory protein interactions, but detailed structural/dynamic characterization of their ensembles remain challenging, both in isolation and when they form dynamic "fuzzy" complexes. Such is the case for mRNA cap-dependent translation initiation, which is regulated by the interaction of the predominantly folded eukaryotic initiation factor 4E (eIF4E) with the intrinsically disordered eIF4E binding proteins (4E-BPs) in a phosphorylation-dependent manner. Single-molecule Förster resonance energy transfer showed that the conformational changes of 4E-BP2 induced by binding to eIF4E are non-uniform along the sequence; while a central region containing both motifs that bind to eIF4E expands and becomes stiffer, the C-terminal region is less affected. Fluorescence anisotropy decay revealed a non-uniform segmental flexibility around six different labeling sites along the chain. Dynamic quenching of these fluorescent probes by intrinsic aromatic residues measured via fluorescence correlation spectroscopy report on transient intra- and inter-molecular contacts on nanosecond-to-microsecond timescales. Upon hyperphosphorylation, which induces folding of ∼40 residues in 4E-BP2, the quenching rates decreased at most labeling sites. The chain dynamics around sites in the C-terminal region far away from the two binding motifs significantly increased upon binding to eIF4E, suggesting that this region is also involved in the highly dynamic 4E-BP2:eIF4E complex. Our time-resolved fluorescence data paint a sequence-level rigidity map of three states of 4E-BP2 differing in phosphorylation or binding status and distinguish regions that form contacts with eIF4E. This study adds complementary structural and dynamics information to recent studies of 4E-BP2, and it constitutes an important step toward a mechanistic understanding of this important IDP via integrative modeling.


Subject(s)
Eukaryotic Initiation Factor-4E , Intrinsically Disordered Proteins , Eukaryotic Initiation Factor-4E/chemistry , Eukaryotic Initiation Factor-4E/genetics , Eukaryotic Initiation Factor-4E/metabolism , Eukaryotic Initiation Factors/chemistry , Eukaryotic Initiation Factors/genetics , Eukaryotic Initiation Factors/metabolism , Intrinsically Disordered Proteins/chemistry , Phosphorylation , Protein Binding
11.
Nanomaterials (Basel) ; 12(4)2022 Feb 17.
Article in English | MEDLINE | ID: mdl-35214997

ABSTRACT

A series of reconfigurable compact photonic arbitrary power splitters are proposed based on the hybrid structure of silicon and Ge2Sb2Se4Te1 (GSST), which is a new kind of non-volatile optical phase change material (O-PCM) with low absorption. Our pixelated meta-hybrid has an extremely small photonic integrated circuit (PIC) footprint with a size comparable to that of the most advanced electronic integrated circuits (EICs). The power-split ratio can be reconfigured in a completely digital manner through the amorphous and crystalline switching of the GSST material, which only coated less than one-fifth of the pattern allocation area. The target power-split ratio between the output channels can be arbitrarily reconfigured digitally with high precision and in the valuable C-band (1530-1560 nm) based on the analysis of three-dimensional finite-difference time-domain. The 1 × 2, 1 × 3, and 1 × 4 splitting configurations were all investigated with a variety of power-split ratios for each case, and the corresponding true value tables of GSST distribution are given. These non-volatile hybrid photonic splitters offer the advantages of an extremely small footprint and non-volatile digital programmability, which are favorable to the truly optoelectronic fusion chip.

12.
Opt Express ; 28(20): 29306-29317, 2020 Sep 28.
Article in English | MEDLINE | ID: mdl-33114833

ABSTRACT

Metasurfaces, the two-dimensional artificial metamaterials, have attracted intensive attention due to their abnormal ability to manipulate the electromagnetic wave. Although there have been considerable efforts to design and fabricate beam steering devices, continuously tunable devices with a uniform bias-voltage have not been achieved. Finding new ways to realize more convenient and simpler wavefront modulation of light still requires research efforts. In this article, a series of novel reflective metasurfaces are proposed to continuously modulate the wavefront of terahertz light by uniformly adjusting the bias-voltage. By introducing the innovation of nonuniform periodic structures, we realize the gradient distribution of the reflected light phase-changing-rate which is the velocity of phase changing with Fermi energy. Based on strict phase distribution design scheme, a beam scanner and a variable-focus reflective metalens are both demonstrated successfully. Furthermore, dynamic and continuous control of either the beam azimuth of beam scanner or the focal length of metalens can be achieved by uniformly tuning the Fermi energy of graphene. Our work provides a potentially efficient method for the development and simplification of the adjustable wavefront controlling devices.

13.
Nat Commun ; 11(1): 3146, 2020 06 19.
Article in English | MEDLINE | ID: mdl-32561718

ABSTRACT

Phosphorylation of intrinsically disordered eIF4E binding proteins (4E-BPs) regulates cap-dependent translation by weakening their ability to compete with eIF4G for eIF4E binding within the translation initiation complex. We previously showed that phosphorylation of T37 and T46 in 4E-BP2 induces folding of a four-stranded beta-fold domain, partially sequestering the canonical eIF4E-binding helix. The C-terminal intrinsically disordered region (C-IDR), remaining disordered after phosphorylation, contains the secondary eIF4E-binding site and three other phospho-sites, whose mechanisms in inhibiting binding are not understood. Here we report that the domain is non-cooperatively folded, with exchange between beta strands and helical conformations. C-IDR phosphorylation shifts the conformational equilibrium, controlling access to eIF4E binding sites. The hairpin turns formed by pT37/pT46 are remarkably stable and function as transplantable units for phospho-regulation of stability. These results demonstrate how non-cooperative folding and conformational exchange leads to graded inhibition of 4E-BP2:eIF4E binding, shifting 4E-BP2 into an eIF4E binding-incompatible conformation and regulating translation initiation.


Subject(s)
Eukaryotic Initiation Factor-4E/metabolism , Intrinsically Disordered Proteins/metabolism , Protein Biosynthesis/physiology , RNA Caps/metabolism , Computational Biology , Eukaryotic Initiation Factor-4E/genetics , Intrinsically Disordered Proteins/genetics , Mutagenesis, Site-Directed , Nuclear Magnetic Resonance, Biomolecular , Phosphorylation/physiology , Protein Binding/genetics , Protein Conformation, alpha-Helical/genetics , Protein Conformation, beta-Strand/genetics , Protein Folding , Protein Processing, Post-Translational/physiology
14.
Opt Express ; 28(7): 9552-9562, 2020 Mar 30.
Article in English | MEDLINE | ID: mdl-32225561

ABSTRACT

Data exchange between different data channels can offer more flexible and advanced functions for many optical networks. In this paper, we propose a switchable and reconfigurable data exchange device for arbitrary two optical mode channels based on three-waveguide-coupling (TWC) switches in mode-division multiplexing (MDM) networks. The working principle of the TWC switches is numerically analyzed using the coupled supermode theory. As a proof of concept, switchable data exchange between arbitrary two mode channels among the first three-order quasi-transverse electric modes is experimentally demonstrated successfully. The insertion losses of the device are less than 5.6 dB, including the coupling loss of the multiplexer and demultiplexer, while the mode crosstalk is less than -13.0 dB for all functions. The proposed device is expected to offer more flexibility to on-chip MDM networks due to its low loss, low crosstalk and good scalability.

15.
Sensors (Basel) ; 20(6)2020 Mar 16.
Article in English | MEDLINE | ID: mdl-32188050

ABSTRACT

In this study, a graphene and optical microfiber coupler (OMC) integrated device (GOMC) was proposed and fabricated. After its characteristic analysis and testing, it was applied to the development of adjustable multi-wavelength fiber lasers. By integrating the OMC with graphene, the polarization dependence of OMC was enhanced. Meanwhile, the novel GOMC was given the capabilities of filtering, coupling, beam splitting, and polarization correlation. When the GOMC was integrated as a filter and beam splitter into the ring cavity of the fiber laser, the proposed GOMC-based fiber laser could achieve single-wavelength and multi-wavelength regulated output. The laser had a 3 dB linewidth of less than 30 pm, a signal-to-noise ratio of approximately 40 dB, and an output power fluctuation of less than 1 dB. The GOMC could also be used for the development of functional devices, such as adjustable mode lockers and mode coupling selectors, which provide an excellent experimental platform for new fiber lasers and the research of multi-dimensional light-field manipulation.

16.
Biochim Biophys Acta Biomembr ; 1859(7): 1242-1253, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28392350

ABSTRACT

Nonspecific interactions between lipids and fluorophores can alter the outcomes of single-molecule spectroscopy of membrane proteins in live cells, liposomes or lipid nanodiscs and of cytosolic proteins encapsulated in liposomes or tethered to supported lipid bilayers. To gain insight into these effects, we examined interactions between 9 dyes that are commonly used as labels for single-molecule fluorescence (SMF) and 6 standard lipids including cationic, zwitterionic and anionic types. The diffusion coefficients of dyes in the absence and presence of set amounts of lipid vesicles were measured by fluorescence correlation spectroscopy (FCS). The partition coefficients and the free energies of partitioning for different fluorophore-lipid pairs were obtained by global fitting of the titration FCS curves. Lipids with different charges, head groups and degrees of chain saturation were investigated, and interactions with dyes are discussed in terms of hydrophobic, electrostatic and steric contributions. Fluorescence imaging of individual fluorophores adsorbed on supported lipid bilayers provides visualization and additional quantification of the strength of dye-lipid interaction in the context of single-molecule measurements. By dissecting fluorophore-lipid interactions, our study provides new insights into setting up single-molecule fluorescence spectroscopy experiments with minimal interference from interactions between fluorescent labels and lipids in the environment.


Subject(s)
Fluorescent Dyes/chemistry , Lipids/chemistry , Hydrophobic and Hydrophilic Interactions , Proteins/chemistry , Spectrometry, Fluorescence , Static Electricity
17.
Langmuir ; 32(42): 10870-10880, 2016 Oct 25.
Article in English | MEDLINE | ID: mdl-27564412

ABSTRACT

Nanotechnology provides a promising platform for drug-delivery in medicine. Nanostructured materials can be designed with desired superparamagnetic or fluorescent properties in conjunction with biochemically functionalized moieties (i.e., antibodies, peptides, and small molecules) to actively bind to target sites. These multifunctional properties make them suitable agents for multimodal imaging, diagnosis, and therapy. Perfluorohexane nanoemulsions (PFH-NEs) are novel drug-delivery vehicles and contrast agents for ultrasound and photoacoustic imaging of cancer in vivo, offering higher spatial resolution and deeper penetration of tissue when compared to conventional optical techniques. Compared to other theranostic agents, our PFH-NEs are one of the smallest of their kind (<100 nm), exhibit minimal aggregation, long-term stability at physiological conditions, and provide a noninvasive cancer imaging and therapy alternative for patients. Here, we show, using high-resolution imaging and correlative techniques, that our PFH-NEs, when in tandem with silica-coated gold nanoparticles (scAuNPs), can be used as a drug-loaded therapeutic via endocytosis and as a multimodal imaging agent for photoacoustic, ultrasound, and fluorescence imaging of tumor growth.

18.
J Am Chem Soc ; 138(36): 11583-98, 2016 09 14.
Article in English | MEDLINE | ID: mdl-27494760

ABSTRACT

G protein-coupled receptors constitute the largest family of transmembrane signaling proteins and the largest pool of drug targets, yet their mechanism of action remains obscure. That uncertainty relates to unresolved questions regarding the supramolecular nature of the signaling complex formed by receptor and G protein. We therefore have characterized the oligomeric status of eGFP-tagged M2 muscarinic receptor (M2R) and Gi1 by single-particle photobleaching of immobilized complexes. The method was calibrated with multiplexed controls comprising 1-4 copies of fused eGFP. The photobleaching patterns of eGFP-M2R were indicative of a tetramer and unaffected by muscarinic ligands; those of eGFP-Gi1 were indicative of a hexamer and unaffected by GTPγS. A complex of M2R and Gi1 was tetrameric in both, and activation by a full agonist plus GTPγS reduced the oligomeric size of Gi1 without affecting that of the receptor. A similar reduction was observed upon activation of eGFP-Gαi1 by the receptor-mimic mastoparan plus GTPγS, and constitutively active eGFP-Gαi1 was predominantly dimeric. The oligomeric nature of Gi1 in live CHO cells was demonstrated by means of Förster resonance energy transfer and dual-color fluorescence correlation spectroscopy in studies with eGFP- and mCherry-labeled Gαi1; stochastic FRET was ruled out by means of non-interacting pairs. These results suggest that the complex between M2R and holo-Gi1 is an octamer comprising four copies of each, and that activation is accompanied by a decrease in the oligomeric size of Gi1. The structural feasibility of such a complex was demonstrated in molecular dynamics simulations.


Subject(s)
GTP-Binding Protein alpha Subunits, Gi-Go/chemistry , Molecular Dynamics Simulation , Receptor, Muscarinic M2/chemistry , Animals , CHO Cells , Cricetulus , GTP-Binding Protein alpha Subunits, Gi-Go/metabolism , Humans , Protein Multimerization , Protein Structure, Quaternary , Receptor, Muscarinic M2/metabolism
19.
Biophys J ; 110(7): 1510-1522, 2016 Apr 12.
Article in English | MEDLINE | ID: mdl-27074677

ABSTRACT

Conformational states of the metastable drkN SH3 domain were characterized using single-molecule fluorescence techniques. Under nondenaturing conditions, two Förster resonance energy transfer (FRET) populations were observed that corresponded to a folded and an unfolded state. FRET-estimated radii of gyration and hydrodynamic radii estimated by fluorescence correlation spectroscopy of the two coexisting conformations are in agreement with previous ensemble x-ray scattering and NMR measurements. Surprisingly, when exposed to high concentrations of urea and GdmCl denaturants, the protein still exhibits two distinct FRET populations. The dominant conformation is expanded, showing a low FRET efficiency, consistent with the expected behavior of a random chain with excluded volume. However, approximately one-third of the drkN SH3 conformations showed high, nearly 100%, FRET efficiency, which is shown to correspond to denaturation-induced looped conformations that remain stable on a timescale of at least 100 µs. These loops may contain interconverting conformations that are more globally collapsed, hairpin-like, or circular, giving rise to the observed heterogeneous broadening of this population. Although the underlying mechanism of chain looping remains elusive, FRET experiments in formamide and dimethyl sulfoxide suggest that interactions between hydrophobic groups in the distal regions may play a significant role in the formation of the looped state.


Subject(s)
Fluorescence Resonance Energy Transfer , Models, Molecular , Polymers/chemistry , src Homology Domains , Animals , Drosophila melanogaster/enzymology , Enzyme Stability , Protein Denaturation/drug effects
20.
Biomacromolecules ; 16(3): 1032-9, 2015 Mar 09.
Article in English | MEDLINE | ID: mdl-25665160

ABSTRACT

A lipid bilayer was deposited on a 3 µm diameter polystyrene (PS) bead via hydrophobic anchoring of bicelles containing oxyamine-bearing cholesteric moieties reacting with the aldehyde functionalized bead surface. Discoidal bicelles were formed by mixing dimyristoylphosphatidylcholine (DMPC), dihexanoylphosphatidylcholine (DHPC), dimyristoyltrimethylammonium propane (DMTAP), and the oxyamine-terminated cholesterol derivative, cholest-5-en-3ß-oxy-oct-3,6-oxa-an-8-oxyamine (CHOLOA), in the molar ratio DMPC/DHCP/DMTAP/CHOLOA (1/0.5/0.01/0.05) in water. Upon exposure to aldehyde-bearing PS beads, a stable single lipid bilayer coating rapidly formed at the bead surface. Fluorescence recovery after photobleaching demonstrated that the deposited lipids fused into an encapsulating lipid bilayer. Electrospray ionization mass spectrometry showed that the short chain lipid DHPC was entirely absent from the PS adherent lipid coating. Fluorescence quenching measurements proved that the coating was a single lipid bilayer. The bicelle coating method is thus simple and robust, can be modified to include membrane-associated species, and can be adapted to coat any number of different surfaces.


Subject(s)
Lipid Bilayers/chemistry , Polystyrenes/chemistry , Dimyristoylphosphatidylcholine/chemistry , Fluorescence Recovery After Photobleaching , Micelles , Phospholipid Ethers/chemistry , Surface Properties , Unilamellar Liposomes/chemistry
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