IndraLab

Statements

E2 activates OTUB1. 21 / 21
| 21
reach
"The effect of E2 enzymes in lowering the K M of OTUB1 for substrate suggests that the binding of an E2 partner increases the affinity of OTUB1 for K48 diubiquitin."
32049508
reach
"Since the N-terminal residues of OTUB1 that precede the OTU domain form a proximal ubiquitin binding helix that plays an essential role in E2 inhibition, we asked whether E2 stimulation of OTUB1 activity similarly requires the OTUB1 N-terminus."
23955022
reach
"No difference in stimulation was observed in cleavage of di-, tri- and tetraubiquitin substrates (XREF_SUPPLEMENTARY), indicating that E2 stimulation of OTUB1 DUB activity is independent of polyubiquitin chain length."
23955022
reach
"Importantly, the fact that a substantial proportion of the E2 partners of OTUB1 are uncharged is consistent with a role for E2 stimulation of OTUB1 in the cell."
23955022
reach
"Our structural and biochemical studies showed how E2 enzymes stimulate the DUB activity of OTUB1."
23955022
reach
"These K d values are comparable to the K M values of OTUB1 in the presence of these E2 partners (XREF_FIG), indicating that all E2 enzymes stimulate OTUB1 by increasing the affinity of the DUB for its K48 diubiquitin substrate."
32049508
reach
"The same E2 enzymes, when uncharged, can stimulate the DUB activity of OTUB1 in vitro, although the importance of OTUB1 stimulation in vivo remains unclear."
32049508
reach
"EC 50 s for E2 stimulation of OTUB1 match intracellular concentrations."
32049508
reach
"Since only uncharged E2 stimulates OTUB1 DUB activity while charged E2 ~ Ub represses OTUB1 under normal cellular concentrations of free ubiquitin, the relative proportion of charged versus uncharged E2s must be critical determinants of whether OTUB1-E2 complexes function as active DUBs in the cell."
23955022
reach
"Thus, the same E2 enzymes that are inhibited by OTUB1 when the E2 is charged with ubiquitin can stimulate OTUB1 when uncharged."
32049508
reach
"For the DUB, we used human OTUB1, which selectively cleaves K48 Ub-Ub linkages 33 and is activated allosterically by certain E2 enzymes 34."
31308549
reach
"We have shown here that a subset of E2 enzymes markedly stimulate OTUB1 cleavage of Lys48 linked polyubiquitin (XREF_FIG) and that the same set of OTUB1-E2 interactions are required for both OTUB1 non catalytic inhibition of E2 enzymes and E2 stimulation of OTUB1 XREF_BIBR, XREF_BIBR."
23955022
reach
"These studies were performed in the presence of OTUB1-stimulating Ubiquitin-conjugating enzyme E2 D1 (UBE2D1), an E2 ubiquitin ligase that engages in a complex with OTUB1 to stimulate OTUB1 activity."
35210618
reach
"Only the uncharged E2 stimulates OTUB1 DUB activity at physiological concentrations of ubiquitin 37, whereas the charged E2 ~ Ub drives formation of a repressed complex with OTUB1 XREF_BIBR, XREF_BIBR, XREF_BIBR."
32049508
reach
"The E2 increased the affinity of OTUB1 for K48 diubiquitin, as reflected in a decrease in K d from 84 muM with no E2 to 12 muM in the presence of UBE2D1, 13.2 muM in the presence of UBE2D3, and 22.3 muM in the presence of UBE2N."
32049508
reach
"Of the E2 enzymes that stimulate OTUB1 (XREF_FIG), UBCH5C (UBE2E3) and UBC13 (UBE2N) have been shown to be the most abundant and in molar excess over OTUB1 20."
23955022
reach
"The same E2 enzymes, when uncharged, can stimulate the DUB activity of OTUB1 in vitro, although the importance of OTUB1 stimulation in vivo remains unclear."
32049508
reach
"Our quantitative study of OTUB1 stimulation by its E2 partners provides a basis for further investigating the relative contribution of these E2 enzymes to OTUB1 DUB activity in cells."
32049508
reach
"E2 enzymes stimulate OTUB1 Lys48 deubiquitinating activity."
23955022
reach
"We show that both stimulation of OTUB1 by E2 enzymes and noncatalytic inhibition of E2 enzymes by OTUB1 occur at physiologically relevant concentrations of both partners."
32049508
reach
"We show that both stimulation of OTUB1 by E2 enzymes and noncatalytic inhibition of E2 enzymes by OTUB1 occur at physiologically relevant concentrations of both partners."
32049508