Quantification of protein secondary structure by (13)C solid-state NMR.

High-resolution (13)C solid-state NMR stands out as one of the most promising techniques to solve the structure of insoluble proteins featuring biological and technological importance. The simplest nuclear magnetic resonance (NMR) spectroscopy method to quantify the secondary structure of proteins uses the areas of carbonyl and alpha carbon peaks. The quantification obtained by fitting procedures depends on the assignment of the peaks to the structure, type of line shape, number of peaks to be used, and other parameters that are set by the operator. In this paper, we demonstrate that the analysis of (13)C NMR spectra by a pattern recognition method-based on the singular value decomposition (SVD) regression, which does not depend on the operator-shows higher correlation coefficients for α-helix and ß-sheet (0.96 and 0.91, respectively) than Fourier transform infrared spectroscopy (FTIR) method. Therefore, the use of (13)C solid-state NMR spectra and SVD is a simple and reliable method for quantifying the secondary structures of insoluble proteins in solid-state.

Gadolinium(III) Complexes with N-Alkyl-N-methylglucamine Surfactants Incorporated into Liposomes as Potential MRI Contrast Agents.

Complexes of gadolinium(III) with N-octanoyl-N-methylglucamine (L8) and N-decanoyl-N-methylglucamine (L10) with 1 : 2 stoichiometry were synthesized and characterized by elemental analysis, electrospray ionization-tandem mass spectrometry (ESI-MS), infrared (IR) spectroscopy, and molar conductivity measurements. The transverse (r 2) and longitudinal (r 1) relaxivity protons were measured at 20 MHz and compared with those of the commercial contrasts. These complexes were incorporated in liposomes, resulting in the increase of the vesicle zeta potential. Both the free and liposome-incorporated gadolinium complexes showed high relaxation effectiveness, compared to commercial contrast agent gadopentetate dimeglumine (Magnevist). The high relaxivity of these complexes was attributed to the molecular rotation that occurs more slowly, because of the elevated molecular weight and incorporation in liposomes. The results establish that these paramagnetic complexes are highly potent contrast agents, making them excellent candidates for various applications in molecular MR imaging.

On resonance phase alternated CWFP sequences for rapid and simultaneous measurement of relaxation times.

T1 and T2 relaxation times have been frequently used as probes for physical-chemical properties in several time-domain NMR applications (TD-NMR) such as food, polymers and petroleum industries. T2 measurements are usually achieved using the traditional Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence because it is a fast and robust method. On the other hand, the traditional methods for determining T1, i.e., inversion and saturation recovery, are time-consuming, driving several authors to develop rapid 1D and 2D methods to obtain T1 and T2 or T1/T2 ratio. However, these methods usually require sophisticated processing and/or high signal to noise ratio (SNR). This led us to develop simple methods for rapid and simultaneous determination of T1 and T2 using Continuous Wave Free Precession (CWFP) and Carr-Purcell Continuous Wave Free Precession (CP-CWFP) pulse sequences. Nevertheless, a drawback of these sequences is that they require specific adjustment of the frequency offset or the time interval between pulses (Tp). In this paper we present an alternative form of these sequences, named CWFPx-x, CP-CWFPx-x, where a train of π/2 pulses with phases alternated by π enable performing the experiments on-resonance and independently of Tp, when Tp

Use of Carr-Purcell pulse sequence with low refocusing flip angle to measure T1 and T2 in a single experiment.

The Carr-Purcell pulse sequence, with low refocusing flip angle, produces echoes midway between refocusing pulses that decay to a minimum value dependent on T(2). When the refocusing flip angle was π/2 (CP(90)) and τ>T(2), the signal after the minimum value, increased to reach a steady-state free precession regime (SSFP), composed of a free induction decay signal after each pulse and an echo, before the next pulse. When τ

Qualitative analysis by online nuclear magnetic resonance using Carr-Purcell-Meiboom-Gill sequence with low refocusing flip angles.

The Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence has been used in many applications of magnetic resonance imaging (MRI) and low-resolution NMR (LRNMR) spectroscopy. Recently, CPMG was used in online LRNMR measurements that use long RF pulse trains, causing an increase in probe temperature and, therefore, tuning and matching maladjustments. To minimize this problem, the use of a low-power CPMG sequence based on low refocusing pulse flip angles (LRFA) was studied experimentally and theoretically. This approach has been used in several MRI protocols to reduce incident RF power and meet the specific absorption rate. The results for CPMG with LRFA of 3π/4 (CPMG(135)), π/2 (CPMG(90)) and π/4 (CPMG(45)) were compared with conventional CPMG with refocusing π pulses. For a homogeneous field, with linewidth equal to Δυ=15 Hz, the refocusing flip angles can be as low as π/4 to obtain the transverse relaxation time (T(2)) value with errors below 5%. For a less homogeneous magnetic field, Δυ=100 Hz, the choice of the LRFA has to take into account the reduction in the intensity of the CPMG signal and the increase in the time constant of the CPMG decay that also becomes dependent on longitudinal relaxation time (T(1)). We have compared the T(2) values measured by conventional CPMG and CPMG(90) for 30 oilseed species, and a good correlation coefficient, r=0.98, was obtained. Therefore, for oilseeds, the T(2) measurements performed with π/2 refocusing pulses (CPMG(90)), with the same pulse width of conventional CPMG, use only 25% of the RF power. This reduces the heating problem in the probe and reduces the power deposition in the samples.