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. Author manuscript; available in PMC: 2016 Jun 27.
Published in final edited form as: Chembiochem. 2016 Mar 21;17(9):816–820. doi: 10.1002/cbic.201600017

Table 1.

Kinetic characterization of DUBs that are used in Ub chain restriction analysis[7] for the diUb FRET pairs 17a–g.[a]

DUB Linkage KM [μM] kcat [S−1] kcat/KM [M−1 S−1]
AMSH Lys63   45.4 0.0027            59
AMSH*[b] Lys63   2.4 0.17      70048
Cezanne Lys11   19.4 1.5      78818
OTUB1 Lys48 ⪢50     n.d.
OTUB1*[b] Lys48   38.6 0.66      17158
OTUD1 Lys63 ⪢50     n.d.        4020
OTUD3 Lys6 ⪢50     n.d.
Lys11  52.4 0.0085          162
Lys63  57.9 0.0061          105
TRABID Lys29   40.1 0.053        1317
Lys33   19.6 0.028        1431
Lys63   54.0 0.034          627
OTUD2 Lys11   87.9 4.4      50253
Lys27 ⪢50     n.d.
Lys29 ⪢50     n.d.
vOTU Lys6   3.6 0.87    242031
Lys11   3.4 0.22      63110
Lys48   8.4 1.0    119952
Lys63   1.4 0.091      66266
USP21 Lys6   2.1 0.12      60633
Lys11   1.4 0.087      62735
Lys33   2.1 0.13      60814
Lys48   1.7 0.067      39181
Lys63   2.7 0.16      60558
[a]

Values in italics were obtained by extrapolation beyond the highest substrate concentration.

[b]

Activated versions of AMSH and OTUB1 were created by fusing them to their activators (STAM and UBE2D2, respectively).[11]