Table 1.

Comparison of the SARS‐CoV‐2 PL^pro kinetic parameters obtained using SPE‐MS with those reported.

	Method	Substrate	${\mathrm{k}}_{\mathrm{cat}}$ /${\mathrm{K}}_{\mathrm{m}}$ [M−1 ⋅ s−1]	${\mathrm{K}}_{\mathrm{m}}$ [μM]
1	SPE‐MS[a]	nsp2/3 substrate 2	64980 ±15256	49.4 ±10.5
2	Fluorescence [5a]	K48 linked Ub2‐Amc	241000±94000	61.23 ±19.76
3	Fluorescence [5a]	ISG15‐Amc	521000±36000	8.50 ±0.54
4	Fluorescence [17b]	RLRGG‐Amc	1840	not reported
5	Fluorescence [14]	Cbz‐RLRGG‐Amc	16000	6.9±1.4
6	Fluorescence [18]	Cbz‐RLRGG‐Amc	not reported	70.92 ±10.15
7	Fluorescence polarization [5b]	ISG15‐TAMRA	30210	not reported
8	Fluorescence polarization [5b]	K48 linked Ub2‐TAMRA	1634	not reported
9	FRET [19]	Dabcyl‐FTLKGGAPTKVTE‐Edans	1074.6±261.9	not reported
10	FRET [8a]	Abz‐FTLKGGAPTKVT−Y(3NO2)R	not reported	1854

[a] Kinetic parameters were determined as described in the Supporting Information (Section 4), Michaelis Menten kinetics are shown in Supporting Figure S5; results are means of three independent runs (n=3; mean ±SD). Abz: 2‐aminobenzoyl. Amc: 7‐amino‐4‐methylcoumarin. Cbz: −C(O)OCH₂Ph. Dabcyl: 4‐([4‐(dimethylamino)phenyl]azo)benzoic acid. Edans: 5‐((2‐aminoethyl)amino)naphthalene‐1‐sulfonic acid. TAMRA: Carboxytetramethylrhodamine.