Spinning, biopolymers

In recent years new NMR techniques offering broad applications in stereochemical analysis have come into use. A prominent example is 2D-NMR (both 2D-resolved and 2D-correlated spectroscopy), which has been extensively applied to biopolymers (149-151). Its use with synthetic polymers has, until now, been limited to but a few cases (152, 153). A further technique, cross-polarization magic-angle spinning spectroscopy (CP-MAS NMR) will be discussed in the section on conformational analysis of solid polymers. 

High-resolution 13C NMR studies have been conducted on intact cuticles from limes, suberized cell walls from potatoes, and insoluble residues that remain after chemical depolymerization treatments of these materials. Identification and quantitation of the major functional moieties in cutin and suberin have been accomplished with cross-polarization magic-angle spinning as well as direct polarization methods. Evidence for polyester crosslinks and details of the interactions among polyester, wax, and cell-wall components have come from a variety of spin-relaxation measurements. Structural models for these protective plant biopolymers have been evaluated in light of the NMR results. 

During the last 15 years, the cross-polarization magic-angle spinning (CPMAS) technique (2) has been used with increasing frequency to provide detailed structural information about solid polymers and biopolymers... 

New techniques for data analysis and improvements in instrumentation have now made it possible to carry out stmctural and conformational studies of biopolymers including proteins, polysaccharides, and nucleic acids. NMR, which may be done on noncrystalline materials in solution, provides a technique complementary to X-ray diffraction, which requires crystals for analysis. One-dimensional NMR, as described to this point, can offer structural data for smaller molecules. But proteins and other biopolymers with large numbers of protons will yield a very crowded spectrum with many overlapping lines. In multidimensional NMR (2-D, 3-D, 4-D), peaks are spread out through two or more axes to improve resolution. The techniques of correlation spectroscopy (COSY), nuclear Overhausser effect spectroscopy (NOESY), and transverse relaxation-optimized spectroscopy (TROSY) depend on the observation that nonequivalent protons interact with each other. By using multiple-pulse techniques, it is possible to perturb one nucleus and observe the effect on the spin states of other nuclei. The availability of powerful computers and Fourier transform (FT) calculations makes it possible to elucidate structures of proteins up to 40,000 daltons in molecular mass and there is future promise for studies on proteins over 100,000... 

Nucleotides - [AMINO AC IDS - L-MONOSODIUM GLUTAMATE (MSG)] (Vol 2) -as antibiotics [ANTIBIOTICS - NUCLEOSIDES AND NUCLEOTIDES] (Vol 3) -catabolism of [MINERALNUTRIENTS] (Vol 16) -electrodes for [BIOPOLYMERS - ANALYTICAL TECHNIQUES] (Vol 4) -phosphorus nmr [MAGNETIC SPIN RESONANCE] (Vol 15) -as radioactive tracers [RADIOACTIVETRACERS] (Vol 20)... 

The 2,2,6,6-tetramethylpiperidinoxyl radical (TEMPO) was first prepared in 1960 by Lebedev and Kazarnovskii by oxidation of its piperidine precursor.18 The steric hindrance of the NO bond in TEMPO makes it a highly stable radical species, resistant to air and moisture. Paramagnetic TEMPO radicals can be employed as powerful spin probes for elucidating the structure and dynamics of both synthetic and biopolymers (e.g., proteins and DNA) by ESR spectroscopy.19 Unlike solid-phase 1H-NMR where magic angle spinning is required in order to reduce the anisotropic effects in the solid-phase environment, solid-phase ESR spectroscopy can be conducted without specialized equipment. Thus, we conducted comparative ESR studies of various polymers with persistent radical labels, and we also determined rotational correlation times as a function of... 

Information about RNA structure and movement is critical for our understanding of how RNA is able to carry out its multifaceted functions. One spectroscopic technique that has shown great promise to study RNA, as well as other biopolymers, is electron paramagnetic resonance (EPR) spectroscopy, also named electron spin resonance (ESR) spectroscopy. EPR is a magnetic resonance technique that monitors the behaviors of unpaired electrons, and has long been used to study structure and dynamics of biomolecules (see recent reviews by Klug and Feix, 2008 Sowa and Qin, 2008). Structural information can be obtained by distance measurements, that is, by determination of distances between two spin-centers, and is the topic of another chapter in this volume (see Chapter 16 in this volume). 

Since RNA is diamagnetic, EPR studies of RNA require incorporation of unpaired electrons into the biopolymer. Nitroxides in five- or six-membered rings that are flanked by methyl groups are stable organic free radicals that are commonly used for spin-labeling (Fig. 15.1). For a free... 

Zhang, Z., et al. (2008). Rotational dynamics of HIV-1 nucleocapsid protein NCp7 as probed by a spin label attached by peptide synthesis. Biopolymers 89, 1125—1135. 

The main source of conformational information for biopolymers are the easy-to-obtain chemical shifts that can be translated into dihedral restraints. In addition, for fully 13C labeled compounds, proton-driven spin diffusion between carbons [72] can be used to measure quantitatively distances between carbons. The CHHC experiment is the equivalent of the NOESY in solution that measures distances between protons by detecting the resonances of the attached carbons. While both techniques, proton-driven spin diffusion and CHHC experiment [73], allow for some variation in the distance as determined from cross-peak integrals, REDOR [74] experiments in selective labeled compounds measure very accurate distances by direct observation of the oscillation of a signal by the dipolar coupling. While the latter technique provides very accurate distances, it provides only one piece of information per sample. Therefore, the more powerful techniques proton-driven spin diffusion and CHHC have taken over when it comes to structure determination by ss-NMR of fully labeled ligands. 

Especially notable are nitroxide radicals such as 11-1. Molecules of this class are sufficiently stable to survive a variety of chemical reactions, allowing them to be covalently bonded to other molecules (e.g., biopolymers such as proteins and nucleic acids). They can then serve as spin labels, transmitting information (via their EPR spectra) about the molecules to which they are attached. 

We used the crosslinked chitosan fiber (hereafter called ChF) in this experimental study. ChF was fabricated by Fuji Spinning Co., Japan. Fig.l shows the unit molecular structure of chitosan which was transformed from chitin by deacetylation. Chitin is a natural biopolymer which is contained in the shell of arthropods. Chitosan was crosslinked to make an adsorbent with acid, alkaline, and chemical proofs. The fabrication method of ChF was presented elsewhere.[S,6]. 

Despite the fact that it may be very difficult to find spin coupling schemes that correspond to realistic value landscapes and can be directly analyzed, the analogy to spin lattices is of great heuristic value. It provides a straightforward explanation of the existence of well-defined error thresholds that sharpen with increasing chain length v, just as cooperative transitions do in linear biopolymers. 

PEO, other polymers such as PCL [66] and biopolymers such as elastin [ 100], were introduced to similarly aid in spinning feasibility [100, 137]. 

Table 1 Comparison of synthetic and biopolymer solvent compatibilities and post-spinning crosslinkers investigated to date... 

For a long time, magic-angle spinning (MAS A has helped biomolecular NMR applications in cases where slow molecular tumbling or susceptibility effects prohibit high-resolution spectroscopy under static conditions. For example, the benehcial effect of MAS has been observed for structural studies on biopolymers, model membranes,... 

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