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1.
Angew Chem Int Ed Engl ; 62(28): e202301969, 2023 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-37066813

RESUMO

While most nanoproteomics approaches for the analysis of low-input samples are based on bottom-up proteomics workflows, top-down approaches enabling proteoform characterization are still underrepresented. Using mammalian cell proteomes, we established a facile one-pot sample preparation protocol based on protein aggregation on magnetic beads and intact proteoform elution using 40 % formic acid. Performed on a digital microfluidics device, the workflow enabled sensitive analyses of single Caenorhabditis elegans nematodes, thereby increasing the number of proteoform identifications compared to in-tube sample preparation by 46 %. Label-free quantification of single nematodes grown under different conditions allowed to identify changes in the abundance of proteoforms not distinguishable by bottom-up proteomics. The presented workflow will facilitate proteoform-directed analysis on samples of limited availability.


Assuntos
Caenorhabditis elegans , Microfluídica , Animais , Caenorhabditis elegans/metabolismo , Proteoma/análise , Proteômica/métodos , Fenômenos Magnéticos , Mamíferos/metabolismo
2.
Cell Rep ; 35(13): 109278, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34192537

RESUMO

A 5,000-year-old Yersinia pestis genome (RV 2039) is reconstructed from a hunter-fisher-gatherer (5300-5050 cal BP) buried at Rinnukalns, Latvia. RV 2039 is the first in a series of ancient strains that evolved shortly after the split of Y. pestis from its antecessor Y. pseudotuberculosis ∼7,000 years ago. The genomic and phylogenetic characteristics of RV 2039 are consistent with the hypothesis that this very early Y. pestis form was most likely less transmissible and maybe even less virulent than later strains. Our data do not support the scenario of a prehistoric pneumonic plague pandemic, as suggested previously for the Neolithic decline. The geographical and temporal distribution of the few prehistoric Y. pestis cases reported so far is more in agreement with single zoonotic events.


Assuntos
Peste/microbiologia , Yersinia pestis/fisiologia , Humanos , Letônia , Funções Verossimilhança , Filogenia
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