Beneficial effects of Glc-1,6-P2 modulation on mutant phosphomannomutase-2_Proteomics

Secretomes and total cell lysates deriving from -/-PMM1 and +/+PMM1 samples were compared by mass spectrometry-based quantitative proteomic through Stable Isotope Dimethyl Labeling. Two biological replicates were analysed for both secretomes and total cell lysates, with the latter being pre-fractionated in 14 fractions.
More in details, peptides were labeled as “light” and “heavy” when belonging to -/-PMM1 and +/+PMM1 samples, respectively. Thus, a heavy/light (H/L) ratio ˃ 1 indicates how much a peptide (and consequently, a protein) is scarce in the -/-PMM1 sample (light) in respect to the +/+PMM1 (heavy) one and vice versa. Bioinformatic analysis was performed through MaxQuant software to identify and quantify proteome alterations. Protein ratios higher than 1.5 and lower than 0.5 were considered reliable alterations of the proteome due to PMM1 knock-out, when existing in both biological replicates.