REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial strains
XL10-Gold Ultracompetent Cells	Agilent Technologies	200317
BL21(DE3) Competent Cells	Agilent Technologies	200131
Chemicals, peptides, and recombinant proteins
Ulp1 Sumo Protease	This work	Described above
Ampicillin sodium salt	Sigma-Aldrich	A9518
Dithiothreitol (DTT)	Sigma-Aldrich	D9779
Isopropyl-β-D-thiogalactopyranoside (IPTG)	Sigma-Aldrich	I6758
Tris(2-carboxyethyl)phosphine hydrochloride (TCEP)	Sigma-Aldrich	C4706
Super Optimal Broth with catabolite repression (SOC) medium	Sigma-Aldrich	S1797
LB broth powder (Miller)	Sigma-Aldrich	L3522
Super broth powder	Research Products International	S33050
Agar	Sigma-Aldrich	A1296
Dnase I	Sigma-Aldrich	DN25
Magnesium chloride (MgCl2)	Sigma-Aldrich	M8266
Sodium chloride (NaCl)	Sigma-Aldrich	S9888
Trizma base (Tris)	Sigma-Aldrich	T6066
Imidazole	Sigma-Aldrich	I2399
Ethylenediaminetetraacetic acid (EDTA)	Sigma-Aldrich	E9884
Iron(III) chloride hexahydrate (FeCl3 × 6H2O)	Sigma-Aldrich	236489
Zinc sulfate heptahydrate (ZnSO4 × 7H2O)	Sigma-Aldrich	221376
Cobalt(II) chloride hexahydrate (CoCl2 × 6H2O)	Sigma-Aldrich	255599
Sodium molybdate dihydrate (Na2MoO4 × 2H2O)	Sigma-Aldrich	M1003
Copper(II) sulfate pentahydrate (CuSO4 × 5H2O)	Sigma-Aldrich	209198
Boric acid (H3BO3)	Sigma-Aldrich	B0394
Manganese(II) sulfate (MnSO4)	Sigma-Aldrich	M7506
Hydrochloric acid (HCl)	Sigma-Aldrich	XX0628
Sodium phosphate monobasic (NaH2PO4)	Sigma-Aldrich	74092
Potassium phosphate monobasic (KH2PO4)	J.T.Baker	3246-05
Potassium phosphate dibasic (K2HPO4)	J.T.Baker	3252-01
Sodium citrate dihydrate (NaC6H5O7 × 2H2O)	J.T.Baker	3646-01
Magnesium sulfate heptahydrate (MgSO4 × 7H2O)	J.T.Baker	2500-01
MEM Vitamin Solution (100×)	Sigma-Aldrich	M6895
15N-ammonium sulfate (15NH4)2SO4	Sigma-Aldrich	299286
12C D-Glucose	Sigma-Aldrich	G7520
D-Glucose (U-13C6,99 %; 1,2,3,4,5,6,6-D7, 97–98 %)	Cambridge Isotope	CLM-1396-5
Deuterium oxide (2H2O)	CortecNet	CD5251P1000
Nickel(II) sulfate (NiSO4)	Sigma-Aldrich	656895
Critical commercial assays
NucleoSpin Plasmid NoLid	Macherey-Nagel	740499.50
Dabsyl-KTSAVLQ/SGFRKME-EDANS	GenScript Inc	Custom Synthesis
5(2Aminoethylamino)1Napthalene sulfonic acid (EDANS)	MedChemExpress	ICN15474001
Recombinant DNA
pGTM_CoV2_NSP5_004_SUMO	This work – available from Addgene	AddGene ID: 190062
pGTM_CoV2_NSP5_C145A_001_SUMO	This work – available from Addgene	AddGene ID: 192349
pGTM_YR375_SUMO_Protease_001	This work – available from Addgene	AddGene ID: 190063
Other
Sonicator 3000	Misonix	S-3000/R2225
Amicon Ultra Centrifugal Filter Unit (MWCO 10000 Da)	Merck Millipore	UFC90110
5-mL HiTrap Chelating HP	Cytiva	17040901
HiPrep 26/10 Desalting	Cytiva	17508701
HiLoad® 16/60 Superdex® 75	Cytiva	28988946/17104401
Infinite M1000 PRO microplate reader	TECAN	30063849/140100256
Jupiter C4 column	Phenomenex	00F-4167-B0
ESI-Q-TOF spectrometer	Xevo, Waters	N/A
Shigemi 5 mm Symmetrical NMR microtube	Shigemi, INC	BMS-005B
Bruker INOVA 700 MHz NMR spectrometer	Bruker	N/A
EasyXtal 15-well crystallization plate	NeXtal	132006 132006
Protein Crystallization Cabinet	RUMED	PKS-E100
Magnetic Cryo Vials	Molecular Dimensions	MD7-402
SPINE Sample Changer Basket	Molecular Dimensions	MD7-510
Magnetic Mounted LithoLoops (0.20 mm)	Molecular Dimensions	MD7-136
Magnetic Cryo wand	Molecular Dimensions	MD7-411
Software and algorithms
NMRPipe	Delaglio, F. et al. (1995)10	https://www.ibbr.umd.edu/nmrpipe/
POKY	Lee, W. et al. (2021)11	https://sites.google.com/view/pokynmr
NMRFAM-SPARKY	Lee, W. et al. (2015)12	https://nmrfam.wisc.edu/nmrfam-sparky-distribution/
MXCuBE	Gabaldinho, J. et al. (2010)13 Oscarsson, M. et al. (2019)14	https://mxcube.github.io/mxcube/
XDS	Kabsch, W. (2010)15	https://strucbio.biologie.uni-konstanz.de/xdswiki/index.php/Xds
CCP4	Winn, M. D. et al. (2011)16	https://www.ccp4.ac.uk/
AIMLESS	Evans, P. R. and Murshudov, G. N. (2013)17	https://www.ccp4.ac.uk/html/aimless.html
REFMAC	Murshudov, G. N. et al. (2011)18	https://www.ccp4.ac.uk/html/refmac5/description.html
COOT	Emsley P. and Cowtan K. (2004)19	https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
Magellan™ v 7.2		https://lifesciences.tecan.com/software-magellan
GraphPad Prism		https://www.graphpad.com
Microsoft Excel		https://www.microsoft.com/en-us/microsoft-365/excel