2 research outputs found
Construction of a Ca<sup>2+</sup>-Gated Artificial Channel by Fusing Alamethicin with a Calmodulin-Derived Extramembrane Segment
Using native chemical ligation, we constructed a Ca<sup>2+</sup>-gated fusion channel protein consisting of alamethicin and
the C-terminal
domain of calmodulin. At pH 5.4 and in the absence of Ca<sup>2+</sup>, this fusion protein yielded a burst-like channel current with no
discrete channel conductance levels. However, Ca<sup>2+</sup> significantly
lengthened the specific channel open state and increased the mean
channel current, while Mg<sup>2+</sup> produced no significant changes
in the channel current. On the basis of 8-anilinonaphthalene-1-sulfonic
acid (ANS) fluorescent measurement, Ca<sup>2+</sup>-stimulated gating
may be related to an increased surface hydrophobicity of the extramembrane
segment of the fusion protein
<i>J</i>‑Resolved <sup>1</sup>H NMR 1D-Projections for Large-Scale Metabolic Phenotyping Studies: Application to Blood Plasma Analysis
<sup>1</sup>H nuclear magnetic resonance (NMR) spectroscopy-based
metabolic phenotyping is now widely used for large-scale epidemiological
applications. To minimize signal overlap present in 1D <sup>1</sup>H NMR spectra, we have investigated the use of 2D <i>J</i>-resolved (JRES) <sup>1</sup>H NMR spectroscopy for large-scale phenotyping
studies. In particular, we have evaluated the use of the 1D projections
of the 2D JRES spectra (pJRES), which provide single peaks for each
of the <i>J</i>-coupled multiplets, using 705 human plasma
samples from the FGENTCARD cohort. On the basis of the assessment
of several objective analytical criteria (spectral dispersion, attenuation
of macromolecular signals, cross-spectral correlation with GC-MS metabolites,
analytical reproducibility and biomarker discovery potential), we
concluded that the pJRES approach exhibits suitable properties for
implementation in large-scale molecular epidemiology workflows