生物化学技术

Treating the analysis of experimental spectroscopic data as an inverse problem and using regularization techniques to obtain stable pseudoinverse solutions, allows access to previously unavailable level of spectroscopic detail. The data is mapped into an appropriate physic ...

Advances in the field of atomic-level membrane simulations are being driven by continued growth in computing power, improvements in the available potential energy functions for lipids, and new algorithms that implement advanced sampling techniques. These developments are allo ...

31P-and 2H-nuclear magnetic resonance are widely used to study order and dynamics, but also orientation of lipid bilayers at solid interfaces. Herein, the various techniques of orienting lipid bilayers at interfaces, the requirements for acquisition of nuclear magnetic resonance s ...

This chapter describes a method of sample preparation called “the rock and roll method,” which is basically a solvent evaporation technique with controlled manual sample movement during evaporation of solvent from lipid/solvent mixtures that produces well-oriented thick stacks ...

Coherent anti-Stokes Raman scattering microscopy develops rapidly into a powerful technique to image both the chemical composition and physical state in complex samples from biophysics, biology, and the material sciences. This nonlinear vibrational technique increases the s ...

Atomic force microscopy (AFM) can be used to reveal intimate details about the effect of anesthetics on phospholipid bilayers. In AFM, surfaces are probed using a tip revealing lateral structural features at 10–20-nm resolution and height features at 0.5-nm resolution. Additionally, in ...

The concentration of long-chain (14–18 carbons) fatty acids (FA) free in solution (unbound) is difficult to measure directly because of the low aqueous solubility of these common dietary FA. One indirect and convenient way to measure the concentration of unbound FA is a method using the fluores ...

Differential-scanning calorimetry is a thermodynamic technique widely used for studying drug-membrane interactions. This chapter provides practical examples on this topic, highlighting the caution to be taken in analyzing thermal data as well as scientific information that ...

In this chapter, a detailed protocol is given for ion-channel reconstitution in the two most used model membranes: planar bilayers and liposomes. In the planar bilayer section, methods are described for the expression of ion channels in Xenopus laevis oocytes, the isolation of their membran ...

Membrane-spanning proteins perturb the organization and dynamics of the adjacent bilayer lipids. For example, when the hydrophobic length (l) of a bilayer-spanning protein differs from the average thickness (d 0) of the host bilayer, the bilayer thickness will vary locally in the vicini ...

The extrinsic fluorescent probe Laurdan (6-dodecanoyl-2-dimethylamino naphthalene) exhibits extreme sensitivity to the polarity and to the molecular dynamics of the dipoles in its environment. Dipolar relaxation processes are reflected as relatively large spectral shi ...

Nuclear magnetic resonance (NMR) spectroscopy enables determination of membrane protein structures in lipid environments, such as micelles and bilayers. This chapter outlines the steps for membrane-protein structure determination using solution NMR with micelle sample ...

Phase-separated supported lipid bilayers have been widely used to study the phase behavior of multicomponent lipid mixtures. One of the primary advantages of using supported lipid bilayers is that the two-dimensional platform of this model membrane system readily allows lipid-pha ...

Cholesterol is a major component of mammalian cell membranes. It has remarkable effects on the properties of phospholipids bilayers, and is implicated in the lipid raft model. Depending on the membrane composition, cholesterol-containing bilayers can exist either as single phase or as ...

Fluorescence microscopy of model membranes is a powerful tool for identifying the nature and extent of coexisting phases in biologically relevant lipid mixtures. Planar supported bilayers offer the advantage over spherical model membranes in that both overall composition and li ...

Near-field scanning optical microscopy (NSOM) allows optical imaging with a spatial resolution that is significantly better than the diffraction limited resolution achievable with conventional optical microscopy. NSOM has the potential to study the nanoscale organizati ...

Lipid rafts and their related membrane vesicular structures, caveolae, are cholesterol- and sphingolipid-rich microdomains of the plasma membrane that have attracted considerable interest because of their ability to concentrate numerous signaling proteins. Efforts to de ...

Bilayers are the most familiar arrangement of phospholipids. However, even as bilayers, phospholipids can arrange themselves in a variety of morphologies from essentially flat structures found in large liposomes or when adhered to a flat solid support, to the curved structures found in s ...

To develop a novel high-throughput tool for monitoring carbohydrate–protein interactions, carbohydrate or glycoprotein microarrays have been prepared for binding with lectins. The interaction events are marked by attachment of fluorescent dyes and gold nanoparticles. The ...

Glycomics which is the study of saccharides and genes responsible for their formation requires the continuous development of rapid and sensitive methods for the identification of glycan structures. It involves glycoanalysis which relies upon the development of methods for deter ...