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Being membranotropic cationites, these "smart nanoparticles" release the overactivating paramagnetic cations only in response to the metabolic acidic shift.
Mutant PtLic16AM2 was selected and characterized, and showed optimal activity at pH 5.0, corresponding to an acidic shift of 2.0 pH units relative to the wild-type enzyme.
Compared with the predicted isoelectric point (pI) of 6.6 (http://us.expasy.org/tools/pi_tool.html) of VER2, the signal detected from vernalized plants (Figure 3A) showed an acidic shift.
Additional spots consistent with multiple phosphorylated forms of 3UIM ataxin-3 were also observed (slight increase in apparent MW with an acidic shift).
As demonstrated in Figure 1B, there were two spots detected by the hK19 antibody, including one with an acidic shift (labeled "2") that was also detected by the anti-pan pY antibody upon analysis of the K19 WT-transfected cell lysates.
Immobilization of MnP enhanced optimum temperature but caused acidic shift in optimum pH of the enzyme.
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The increase of the pore water conductivity (dominated by the increase of ions concentration in solution) along with a transition to acidic conditions shifts the polarization peak towards higher frequencies.
Usually, after purification, the presence of acidic groups shifts IEP to the lowest pH values [25, 26].
Two examples of changed PTM patterns were directly detectable on our 2D gels via acidic spot shifts (actin and septin).
Actin was found in four different protein spots with molecular weight (MW) higher than the theoretical value and acidic pI shifts, indicating posttranslational modifications.
These acidic pI shifts of the protein spots without apparent change of MW could be caused by post-translational modifications (PTMs) such as acetylation and/or phosphorylation.
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