Trapped ion mobility spectrometry-time-of-flight mass spectrometry (TIMS-TOFMS) has emerged as a tool to study protein conformational states. In TIMS, gas-phase ions are guided across the IM stages by applying direct current (DC) potentials (D1-6), which, however, might induce changes in protein structures through collisional activation. To define conditions for native protein analysis, we evaluated the influence of these DC potentials using the metalloenzyme bovine carbonic anhydrase (BCA) as primary test compound. The variation of DC potentials did not change BCA-ion charge and heme content but affected (relative) charge-state intensities and adduct retention. Constructed extracted-ion mobilograms and corresponding collisional cross-section (CCS) profiles gave useful insights in (alterations of) protein conformational state. For BCA, the D3 and D6 potential (which are applied between the deflection transfer and funnel 1 [F1] and the accumulation exit and the start of the ramp, respectively) had most profound effects, showing multimodal CCS distributions at higher potentials indicating gradual unfolding. The other DC potentials only marginally altered the CCS profiles of BCA. To allow for more general conclusions, five additional proteins of diverse molecular weight and conformational stability were analyzed, and for the main protein charge states, CCS profiles were constructed. Principal component analysis (PCA) of the obtained data showed that D1 and D3 exhibit the highest degree of correlation with the ratio of folded and unfolded protein (F/U) as extracted from the mobilograms obtained per set D potential. The correlation of D6 with F/U and protein charge were similar, and D2, D4, and D5 showed an inverse correlation with F/U but were correlated with protein charge. Although DC boundary values for induced conformational changes appeared protein dependent, a set of DC values could be determined, which assured native analysis of most proteins.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1002/jms.5021 | DOI Listing |
Naunyn Schmiedebergs Arch Pharmacol
January 2025
Department of Microbiology, Hind Institute of Medical Sciences, Mau, Ataria, Sitapur, Uttar Pradesh, India, 261303.
Methicillin-resistant Staphylococcus aureus (MRSA) continues to pose significant challenges in healthcare settings due to its multi-drug resistance (MDR) and virulence. This retrospective study examines the molecular and resistance profiles of MRSA isolates from a tertiary care hospital in Saudi Arabia, providing valuable insights into regional epidemiology. A total of 190 MRSA strains were analysed to assess antimicrobial susceptibility, genetic diversity, and virulence factors.
View Article and Find Full Text PDFMicroorganisms
November 2024
Department of Biochemistry, Genetics and Microbiology, University of Pretoria, Private Bag X20, Hatfield, Pretoria 0028, South Africa.
In the absence of data on the reporting of resistance to antibiotics, we sought to determine which clonal complexes (CCs)/sequence types (STs) circulate in the food chain in Kosovo and to determine their antibiogram profiles to a panel of 18 antibiotics. From a total of 114 isolates, 21 different typical STs were identified by multilocus sequence typing (MLST). Each isolate derived from the food categories was subjected to tests to verify its susceptibility to the selected antibiotics according to the designed Sensititre GPN3F panel.
View Article and Find Full Text PDFInt J Environ Res Public Health
November 2024
Department of Liberal Arts and Science, VCU in Qatar, Doha P.O. Box 1129, Qatar.
The COVID-19 pandemic has highlighted the crucial role of health sector decision-makers in establishing and evaluating effective treatment and prevention policies. To inform sound decisions, it is essential to simultaneously monitor multiple pandemic characteristics, including transmission rates, infection rates, recovery rates (which indicate treatment efficacy), and fatality rates. This study introduces an innovative application of existing methodologies: the Multivariate Exponentially Weighted Moving Average (MEWMA) and Multivariate Cumulative Sum (MCUSUM) control charts (CCs), used for monitoring the parameters of the Susceptible, Exposed, Infected, Recovered, Death, and Vaccination (SEIRDV) model.
View Article and Find Full Text PDFBehav Brain Res
January 2025
Experimental Biology Center, University of Fortaleza, Av. Washington Soares, 1321, Fortaleza, Ceará, Brazil; Laboratory of Bioprospection of Natural Products and Biotechnology, Department of Chemistry, CECITEC/UECE - Center for Education, Science and Technology of the Inhamuns Region. R. Seis, 15 - Bezerra de Sousa, Tauá, Ceará, Brazil. Electronic address:
Mimosa tenuiflora ("jurema-preta") is traditionally used in folk medicine for various diseases. The study investigated the neuropharmacological potential of Mimosa tenuiflora bark fraction (FATEM) in adult zebrafish. This included the acute toxicity (LC50) of FATEM (0.
View Article and Find Full Text PDFPlanta
January 2025
ICAR-National Institute for Plant Biotechnology, New Delhi, 110012, Delhi, India.
Small RNA sequencing analysis in two chickpea genotypes, JG 62 (Fusarium wilt-susceptible) and WR 315 (Fusarium wilt-resistant), under Fusarium wilt stress led to identification of 544 miRNAs which included 406 known and 138 novel miRNAs. A total of 115 miRNAs showed differential expression in both the genotypes across different combinations. A miRNA, Car-miR398 targeted copper chaperone for superoxide dismutase (CCS) that, in turn, regulated superoxide dismutase (SOD) activity during chickpea-Foc interaction.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!