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Diaminoanthraquinone Enhances Alkaloid Ionization in MALDI-MS

ABSTRACT: The alkaloids epinastine, 3-methylxanthine and camptothecin were analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The ionization efficiencies of epinastine and 3-methylxanthine were improved upon the addition of 1,5-diaminoanthraquinone (DAAQ). DAAQ did not show ultraviolet absorbance peaks at wavelengths around 337 nm and 355 nm that are used in conventional MALDI-MS instruments. In addition, the DAAQ ion peak was very weak relative to those of the analytes due to the low absorbance efficiency. These properties of DAAQ are advantageous for the DAAQ-MALDI-MS analysis of alkaloids.

SUBMITTER: Yamagaki T

PROVIDER: S-EPMC9853952 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature

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Project description:Highly specialized cells are fundamental for proper functioning of complex organs. Variations in cell-type specific gene expression and protein composition have been linked to a variety of diseases. Although single cell technologies have emerged as valuable tools to address this cellular heterogeneity, a majority of these workflows lack sufficient in situ resolution for functional classification of cells and are associated with extremely long analysis time, especially when it comes to in situ proteomics. In addition, lack of understanding of single cell dynamics within their native environment limits our ability to explore the altered physiology in disease development. This limitation is particularly relevant in the mammalian brain, where different cell types perform unique functions and exhibit varying sensitivities to insults. The hippocampus, a brain region crucial for learning and memory, is of particular interest due to its obvious involvement in various neurological disorders. Here, we present a combination of experimental and data integration approaches for investigation of cellular heterogeneity and functional disposition within the mouse brain hippocampus using MALDI Imaging mass spectrometry (MALDI-IMS) and shotgun proteomics (LC-MS/MS) coupled with laser-capture microdissection (LCM) along with spatial transcriptomics. Within the dentate gyrus granule cells we identified two proteomically distinct cellular subpopulations that are characterized by a substantial number of discriminative proteins. These cellular clusters contribute to the overall functionality of the dentate gyrus by regulating redox homeostasis, mitochondrial organization, RNA processing, and microtubule organization. Importantly, most of the identified proteins matched their transcripts, verifying the in situ protein identification and supporting their functional analyses. By combining high-throughput spatial proteomics with transcriptomics, our approach enables reliable near-single-cell scale identification of proteins and profiling of inter-cellular heterogeneity within similar cell-types in tissues. This methodology has the potential to be applied to different biological conditions and tissues, providing a deeper understanding of cellular subpopulations in situ.

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Diaminoanthraquinone Enhances Alkaloid Ionization in MALDI-MS

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