Journal
JOURNAL OF PROTEOME RESEARCH
Volume 10, Issue 9, Pages 4165-4176Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr200328f
Keywords
metabolomics; NMR; MS; metabolic profiling; B lymphocytes; plasma cells; protein secretion; immunoglobulin; footprinting
Categories
Funding
- Fondazione Cariplo
- AIRC [9965]
- Fondazione Telethon
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When small B lymphocytes bind antigen in the context of suitable signals, a profound geno-proteomic metamorphosis is activated that generates antibody-secreting cells. To study the metabolic changes associated with this differentiation program, we compared the exometabolome of differentiating murine B lymphoma cells and primary B cells by monodimensional proton nuclear magnetic resonance spectroscopy and mass spectrometry coupled to liquid chromatography. Principal component analysis, a multivariate statistical analysis, highlighted metabolic hallmarks of the sequential differentiation phases discriminating between the proliferation and antibody secreting phases and revealing novel metabolic pathways. During proliferation, lactate production increased together with consumption of essential amino acids; massive Ig secretion was paralleled by alanine and glutamate production, glutamine being used as carbon and energy sources. Notably, ethanol and 5'-methylthioadenosine were produced during the last phase of protein secretion and the proliferative burst, respectively. Our metabolomics results are in agreement with previous genoproteomics studies. Thus, metabolic profiling of extracellular medium is a useful tool to characterize the functional state of differentiating B cells and to identify novel underlying metabolic pathways.
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