This study presents a new chemical cross-linking mass spectrometry (MS) method in combination with electrochemistry and isotope labeling strategy for probing both protein three-dimensional (3D) structures and conformational changes. For the former purpose, the target protein/protein complex is cross-linked with equal mole of premixed light and heavy isotope labeled cross-linkers carrying electrochemically reducible disulfide bonds (i.e., DSP-d0 and DSP-d8 in this study, DSP = dithiobis[succinimidyl propionate]), digested and then electrochemically reduced followed with online MS analysis. Cross-links can be quickly identified because of their reduced intensities upon electrolysis and the presence of doublet isotopic peak characteristics. In addition, electroreduction converts cross-links into linear peptides, facilitating MS/MS analysis to gain increased information about their sequences and modification sites. For the latter purpose of probing protein conformational changes, an altered procedure is adopted, in which the protein in two different conformations is cross-linked using DSP-d0 and DSP-d8 separately, and then the two protein samples are mixed in 1:1 molar ratio. The merged sample is subjected to digestion and electrochemical mass spectrometric analysis. In such a comparative cross-linking experiment, cross-links could still be rapidly recognized based on their responses to electrolysis. More importantly, the ion intensity ratios of light and heavy isotope labeled cross-links reveal the conformational changes of the protein, as exemplified by examining the effect of Ca2+ on calmodulin conformation alternation. This new cross-linking MS method is fast and would have high value in structural biology.
In this paper, we give three unbounded conditions under which we are able to solve the nonlinear programming problems in unbounded sets by a homotopy continuation method. In addition, we also discuss their relations. 相似文献
Based on the widely used Gullstrand-Le Grand eye model, the individual human eye model has been established here, which has individual corneal data, anterior chamber depth and the eyeball depth. Furthermore, the foremost thing is that the wavefront aberration calculated from the individual eye model is equal to the eye's wavefront aberration measured with the Hartmann-shack wavefront sensor. There are four main steps to build the model. Firstly, the corneal topography instrument was used to measure the corneal surfaces and depth. And in order to input cornea into the optical model, high-order aspheric surface-Zernike Fringe Sag surface was chosen to fit the corneal surfaces. Secondly, the Hartmann-shack wavefront sensor, which can offer the Zernike polynomials to describe the wavefront aberration, was built to measure the wavefront aberration of the eye. Thirdly, the eye's axial lengths among every part were measured with A-ultrasonic technology. Then the data were input into the optical design software–ZEMAX and the crystalline lens's shapes were optimized with the aberration as the merit function. The individual eye model, which has the same wavefront aberrations with the real eye, is established. 相似文献
A systematic investigation into the influence of the degree of fluorination on the static and dynamic wetting behavior of TiO2‐based nanobelt (TNB) particles with various liquids is described. The effect of the degree of fluorination and the surface tension of the liquid on the occurrence and stability of liquid marbles, foams or dispersions are studied and the wetting behavior and arrangement of particles at the air–liquid surface are observed. Using contact angle (θ) measurements, the relation between the type of particle‐stabilized material and θ is established. For liquids of relatively high tension like water or formamide which do not wet the fluorinated particles, a powder‐like material (marble) is formed. For polar oils of intermediate tension (35–50 mN m?1), which partially wet the fluorinated particles, stable air‐in‐oil foams can be prepared in which particles form a close‐packed layer enveloping air bubbles. Liquids of relatively low tension, e.g., ethanol or polydimethylsiloxane, wet the particles forming a uniform dispersion and partial sedimentation. By contrast, the as‐prepared hydrophilic TNB particles are rapidly wetted by all the liquids as expected due to their high surface energy. The stable cross‐stacked TNB particles with fluoroalkylsilane (FAS) modification could be a versatile platform in a wide range of applications, especially for fluidic devices (e.g., biofluids, gas sensing, and lab‐on‐a‐chip devices). In a proof‐of‐concept study, the oil–water separation performance of fabrics with chemically stable TNB/FAS coating and the liquid isolation by a TNB/FAS shell for highly sensitive gas sensing or reagent assays are investigated. 相似文献
In this paper the fabrication, characterization, and thermal assisted bonding of two photosensitive silicate glasses is presented, as part of a wider process aiming to obtain a planar waveguide lossless splitter for telecom applications. Two glass compositions are investigated: an Er–Yb doped silicate glass, which amplifies the signals when pumped at 980 μm and a matchable passive glass, which is designed to host the waveguide splitter. The two parts were bonded successfully using direct bonding technique at 605 ± 20 °C for 240 min under a pressure of 28 kPa. Vickers hardness measurements were made on the bulk glasses and at the bonded interface, which showed no cracks propagating along the interface. 相似文献