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CSF proteome mapping: Human cerebrospinal fluid LC-MS/MS, Part 3

ABSTRACT: 3 mL CSF from a pool of 21 neurologically healthy donors was in-solution trypsin digested followed by Solid-phase enrichment of N-linked glycopeptides (SPEG) using hydrazide modified beads. Glycopeptides were further fractionated into 20 fractions by mixed-mode reversed phase anion exchange (MM(RP-AX) using a Promix MP column. Each fraction was analyzed separately by LC-MS/MS on an Orbitrap Velos Pro mass spectrometer. Resulting data was searched using SeachGui, summarized in PeptideShaker and exported to the CSF-PR.

INSTRUMENT(S): LTQ Orbitrap, MS:1000031

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Astrid Guldbrandsen

PROVIDER: MSV000098343 | MassIVE | Thu Jun 26 17:11:00 BST 2025

SECONDARY ACCESSION(S): PXD000653

REPOSITORIES: MassIVE

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Project description:The speG gene has been reported to regulate polyamine metabolism in Escherichia coli and Shigella, but its role in Salmonella remains unknown. Our preliminary studies have revealed that speG widely affects the transcriptomes of infected in vitro M and Caco-2 cells and that it is required for the intracellular replication of Salmonella enterica serovar Typhimurium (S. Typhimurium) in HeLa cells. In this study, we demonstrated that speG plays a time-dependent and cell type-independent role in the intracellular replication of S. Typhimurium. Moreover, high-performance liquid chromatography of four major polyamines demonstrated putrescine, spermine, and cadaverine as the leading polyamines in S. Typhimurium. The deletion of speG significantly increased the levels of the three polyamines in intracellular S. Typhimurium, suggesting the inhibitory effect of speG on the biosynthesis of these polyamines. The deletion of speG was associated with elevated levels of these polyamines in the attenuated intracellular replication of S. Typhimurium in host cells, which was subsequently validated by the dose-dependent suppression of intracellular proliferation after the addition of the polyamines. Furthermore, our RNA transcriptome analysis of S. Typhimurium SL1344 and its speG mutant outside and inside Caco-2 cells revealed that speG regulates the genes with documented virulence in flagellar biosynthesis, fimbrial expression, and functions of types III and I secretion systems. speG also affects the expression of genes that have been rarely reported to correlate with polyamine metabolism, including those associated with the periplasmic nitrate reductase system, glucarate metabolism, the phosphotransferase system, cytochromes, and the succinate reductase complex in S. Typhimurium in the mid-log growth phase, as well as those in the ilv–leu and histidine operons of intracellular S. Typhimurium after invasion in Caco-2 cells. In the present study, we characterized the phenotypes and transcriptome effects of speG in S. Typhimurium and reviewed the relevant literature to facilitate a more comprehensive understanding of the key role of speG in the polyamine metabolism and virulence regulation of Salmonella.

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CSF proteome mapping: Human cerebrospinal fluid LC-MS/MS, Part 3

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