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UBE2D2 activates OTUB1. 5 / 8
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"To confirm that the observed stimulation was not assay dependent, we used a gel based assay to verify that UBCH5B stimulates OTUB1 cleavage of native Lys48 linked diubiquitin (XREF_FIG and XREF_SUPPLEMENTARY)."
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"Thes results indicate that UBCH5B stimulates OTUB1 by increasing its affinity for the Lys48 diubiquitin substrate."
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"We assayed the ability of UBCH5B ~ Ub to stimulate OTUB1 (XREF_FIG) and found that both charged and uncharged UBCH5B stimulate OTUB1 to a similar degree."
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"The importance of the OTUB1 N-terminal helix to its ability to be stimulated by UBCH5B, together with the role of these N-terminal residues in forming the proximal ubiquitin binding site of OTUB1 XREF_BIBR, XREF_BIBR, XREF_BIBR, suggested that UBCH5B may increase OTUB1 affinity for Lys48 diubiquitin via the proximal ubiquitin binding site."
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"However, assays of OTUB1 stimulation by UBCH5B or UBCH5B ~ Ub over a range of ubiquitin concentrations (XREF_FIG) (XREF_SUPPLEMENTARY) showed that stimulation by UBCH5B is relatively insensitive to free ubiquitin, but that stimulation by UBCH5B ~ Ub decreases as a function of increasing concentration of free ubiquitin, dropping to low levels below ~ 1 microM free ubiquitin."
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