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Quantitative Proteomic Analysis Reveals Central Metabolic Pathways Affect Norfloxacin Tolerance in Vibrio alginolyticus

ABSTRACT: Antibiotic tolerance often involves reversible metabolic reprogramming. Here, we applied DIA-based quantitative proteomics to profile Vibrio alginolyticus (V. alginolyticus) under norfloxacin (NOR) exposure and observed widespread decreases in protein abundance, consistent with a global metabolic downshift. The most prominent changes were in central carbon and energy metabolism, encompassing the Embden–Meyerhof–Parnas (EMP), pentose phosphate (PPP), and tricarboxylic acid (TCA) pathways, alongside amino acid metabolism and oxidative phosphorylation. . The study utilizes advancedproteomics techniques, including LC-/MS/MS,to uncoverdifferential protein expression patterns.

ORGANISM(S): Vibrio Alginolyticus Nbrc 15630 = Atcc 17749

SUBMITTER: Xiangmin Lin

PROVIDER: PXD072925 | iProX | Sun Jan 11 00:00:00 GMT 2026

REPOSITORIES: iProX

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Project description:Many cells in mammals exist in the state of quiescence, which is characterized by reversible exit from the cell cycle. Quiescent cells are widely reported to exhibit reduced size, nucleotide synthesis, and metabolic activity. Much lower glycolytic rates have been reported in quiescent compared with proliferating lymphocytes. In contrast, we show here that primary human fibroblasts continue to exhibit high metabolic rates when induced into quiescence via contact inhibition. By monitoring isotope labeling through metabolic pathways and quantitatively identifying fluxes from the data, we show that contact-inhibited fibroblasts utilize glucose in all branches of central carbon metabolism at rates similar to those of proliferating cells, with greater overflow flux from the pentose phosphate pathway back to glycolysis. Inhibition of the pentose phosphate pathway resulted in apoptosis preferentially in quiescent fibroblasts. By feeding the cells labeled glutamine, we also detected a “backwards” flux in the tricarboxylic acid cycle from α-ketoglutarate to citrate that was enhanced in contact-inhibited fibroblasts; this flux likely contributes to shuttling of NADPH from the mitochondrion to cytosol for redox defense or fatty acid synthesis. The high metabolic activity of the fibroblasts was directed in part toward breakdown and resynthesis of protein and lipid, and in part toward excretion of extracellular matrix proteins. Thus, reduced metabolic activity is not a hallmark of the quiescent state. Quiescent fibroblasts, relieved of the biosynthetic requirements associated with generating progeny, direct their metabolic activity to preservation of self integrity and alternative functions beneficial to the organism as a whole. mRNAs were analyzed by two color microarray from two separate human neonatal dermal fibroblasts cell lines in proliferating, 7 days contact inhibition, or 14 days contact inhibition. Contact inhibited samples were co-hybridized to proliferating samples as a control, while an additional array co-hybridized the two proliferating samples to analyze reproducibility.

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Quantitative Proteomic Analysis Reveals Central Metabolic Pathways Affect Norfloxacin Tolerance in Vibrio alginolyticus

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