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It therefore appears that DUB activity, rather than a specific DUB, is the crucial determinant in imparting discrimination in this system.
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Furthermore, using structure-based mutants, we identify a triad of residues in the methyllysine binding pocket that imparts discrimination between monomethyllysine and dimethyllysine.
Other purified DUBs (USP7, Ataxin3) could also impart discrimination in this assay, albeit to different extents that correlated with their relative catalytic activities (data not shown).
Use of a purified DUB (the catalytic domain of USP2) in lieu of cytosol also imparted discrimination to the radiolabeled recombinant mCD4-Vpu system in a concentration-dependent manner.
Thus, the radiolabeled recombinant in vitro system seems to lack some feature(s) that imparts substrate discrimination to the crude in vitro and cellular systems (summarized in Figure 4F).
These technical quirks notwithstanding, at least one contributing factor to discrimination imparted by cytosol can be attributed to DUB activity.
What about imparting Catholic values?
You're imparting values.
We propose that discrimination with respect to eventual degradation is imparted by a combination of four parameters.
I impart my knowledge.
Any lessons to impart?
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