Native mass spectrometry has emerged as an important tool for gas-phase structural biology. However, the conformations that a biomolecular ion adopts in the gas phase can differ from those found in solution. Herein, we report a synergistic, native ion mobility-mass spectrometry (IM-MS) and transition metal ion Forster resonance energy transfer (tmFRET)based approach to probe the gas-phase ion structures of a nonstapled peptide (nsp; Ac-CAARAAHAAAHARARA-NH 2 ) and a stapled peptide (sp; Ac-CXARAXHAAAHARARA-NH 2 ). The stapled peptide contains a single hydrocarbon chain connecting the peptide backbone in the i and i + 4 positions via a Grubbs ring-closure metathesis. Fluorescence lifetime measurements indicated that the Cu-bound complexes of carboxyrhodamine 6g (crh6g)-labeled stapled peptide (sp-crh6g) had a shorter donor−acceptor distance (r DA ) than the labeled nonstapled peptide (nsp-crh6g). Experimental collision cross-section (CCS) values were then determined by native IM-MS, which could separate the conformations of Cu-bound complexes of nsp-crh6g and sp-crh6g. Finally, the experimental CCS (i.e., shape) and r DA (i.e., distance) values were used as constraints for computational studies, which unambiguously revealed how a staple reduces the elongation of the peptide ions in the gas phase. This study demonstrates the superiority of combining native IM-MS, tmFRET, and computational studies to investigate the structure of biomolecular ions.
Even populations of clonal cells are heterogeneous, which requires high-throughput analysis methods with singlecell sensitivity.H ere,w ep ropose ar apid, label-free single-cell analytical method based on active capillary dielectric barrier discharge ionization mass spectrometry,w hich can analyze multiple metabolites in single cells at arate of 38 cells/minute. Multiple cell types (HEK-293T,P ANC-1, CFPAC-1, H6c7, HeLa and iBAs) were discriminated successfully.W ef ound evidence for abnormal lipid metabolism in pancreatic cancer cells.W ea lso analyzed gene expression in ac ancer genome atlas dataset and found that the mRNAl evel of ac ritical enzyme of lipid synthesis (ATP citrate lyase,A CLY) was upregulated in human pancreatic ductal adenocarcinoma (PDAC). Moreover,b oth an ACLY chemical inhibitor and asiRNAapproachtargeting ACLY could suppress the viability of PDAC cells.Asignificant reduction in lipid content in treated cells indicates that ACLY could be apotential target for treating pancreatic cancer.
A disposable, instrument-free, height readout paper-based analytical device (HR-PAD) based on a paper strip inkjet-printed with CdTe quantum dots (QDs) was developed for the sensitive speciation analysis of Ag + and silver nanoparticles (AgNPs). When the paper strip is immersed into a sample solution, capillary action draws it through the surface and any Ag + in the solution quenches the fluorescence of CdTe QDs via a cation exchange reaction between Ag + and the CdTe QDs, with the height of the quenched band being proportional to the concentration of Ag + . In contrast, fluorescence quenching cannot be observed when only AgNPs are present in solution. Thus, the concentration of AgNPs can be obtained by subtracting the Ag + content from the total silver determined by the HR-PAD after digestion with HNO 3 . Under optimized conditions, the methodology provides high selectivity, sensitivity, and accuracy for the detection of Ag + or AgNPs in various samples, even at concentrations as low as 0.05 mg L −1 . Precisions of 4.5% and 2.2% RSDs were achieved at concentrations of 1 mg L −1 and 7 mg L −1 of Ag + , respectively. Compared to conventional methods, this approach is inexpensive and user-friendly and eliminates the need for expensive and sophisticated detection instruments. The practicality of the method was demonstrated via the speciation analysis of AgNPs and Ag + in river water and 12 commercial products with satisfactory results.
Single-cell metabolomics is expected to deliver fast and dynamic information on cell function; therefore, it requires rapid analysis of a wide variety of very small quantities of metabolites in living cells. In this work, a hybrid ionization source that combines nanoelectrospray ionization (nanoESI) and dielectric barrier discharge ionization (DBDI) is proposed for single-cell analysis. A capillary with a 1 μm i.d. tip was inserted into cells for sampling and then directly used as the nanoESI source for ionization of polar metabolites. In addition, a DBDI source was employed as a post-ionization source to improve the ionization of apolar metabolites in cells that are not easily ionized by ESI. By increasing the voltage of the DBDI source from 0 to 3.2 kV, the classes of detected metabolites can be shifted from mostly polar to both polar and apolar to mainly apolar. Plant cells (onion) and human cells (PANC-1) were investigated in this study. After optimization, 50 compounds in onion cells and 40 compounds in PANC-1 cells were observed in ESI mode (3.5 kV) and an additional 49 compounds in onion cells and 73 compounds in PANC-1 cells were detected in ESI (3.5 kV)-DBDI (2.6 kV) hybrid mode. This hybrid ionization source improves the coverage, ionization efficiency, and limit of detection of metabolites with different polarities and could potentially contribute to the fast-growing field of single-cell metabolomics.
Rationale Fragrances are organic compounds with pleasant odors that are widely used in every aspect of our daily life; some fragrance ingredients can cause allergic reactions. Hence, the qualitative and quantitative analysis of fragrance allergens can prevent consumers coming into contact with these compounds. In this study, we evaluated the ability of a dielectric barrier discharge ionization (DBDI) source for analyzing allergens that occur in fragrances. Methods A home‐built liquid‐infusion device was used to evaporate the liquid samples. An active capillary plasma ionization source, which is based on a dielectric barrier discharge, was used to ionize the analytes. Mass spectra were acquired in positive ion mode with an LTQ Orbitrap mass spectrometer. Results Seven typical fragrance allergens were analyzed in this study. The limits of detections (LODs) were as low as 0.0001 ppm and a linear dynamic range of 2–3 orders of magnitude was achieved. Allergens in five different perfume products were successfully analyzed and quantified by this method, with analysis times of less than 1 min per sample. Conclusions This work introduces a DBDI‐MS‐based analytical method for detecting and quantifying fragrance allergens. Since DBDI has the advantages of high sensitivity, simple operation and fast analysis time, it is very suitable for the rapid analysis of trace allergens in fragrances, and could easily be used for quality control of consumer products in the cosmetics market.
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