Skip to main content
. 2020 Dec 24;22(1):124. doi: 10.3390/ijms22010124

Figure 2.

Figure 2

Figure 2

Analysis of Tα1-PAS N-acetylation, peptide purity, and hydrodynamic volume. (a) The deconvoluted ESI-MS spectrum of the Tα1-PAS flow-through fraction after the subtractive AEX step (see text) shows a molecular mass of 52,734.56 Da, corresponding to N-acetylated Tα1-PAS (calc. mass: 52,734.56 Da). (b) ESI-MS analysis of the bound and eluted peptide fraction from the same chromatography reveals a molecular mass of 52,692.37 Da, which matches the calculated mass of non-acetylated Tα1-PAS (52,692.54 Da). (c) Reverse-phase chromatography of the flow-through fraction indicates a purity > 96% for N-acetylated Tα1-PAS. (d) Analytical SEC of the finally purified Tα1-PAS in the presence of phosphate-buffered saline (PBS) results in a single peak with an elution volume of 13.5 mL. Comparison with a half-logarithmic calibration curve (inset) indicates an expanded hydrodynamic volume with an apparent molecular size of 557 kDa.