Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene | mKO2 | Thermo Fisher Scientific |
NCBI accession: AB359188 |
GeneArt synthetic construct |
| Strain (Escherichia coli) |
E. coli Tuner(DE3) | EMD-Millipore | ||
| Strain (Mycobacterium smegmatis mc(2) 155) |
Wild-type strain (Msm WT) |
ATCC | ATCC: 700084 | |
| Genetic reagent (Mycobacterium smegmatis mc(2) 155) |
Msm ∆cpa | This paper | Unmarked cpa deletion strain |
|
| Antibody | anti-RpoBEc | BioLegend | BioLegend clone: 8RB13 |
Antibody against beta-subunit of RNA polymerase from E. coli; used at dilution 1:1000 |
| Antibody | anti-CpaMsm | This paper | Rabbit polyclonal antibody against full-length Cpa from M. smegmatis mc(2) 155; produced by BioGenes GmbH; used at dilution 1:40000 |
|
| Recombinant DNA reagent |
pET28a-His6- TEV-RerCpa |
This paper | Plasmid for expression of rhodococcal Cpa |
|
| Recombinant DNA reagent |
pET28a-His6- TEV-MsmCpa |
This paper | Plasmid for expression of mycobacterial Cpa |
|
| Recombinant DNA reagent |
pET28a-His6-TEV-Rer
Cpa_D312N_E566Q |
This paper | Plasmid for expression of double Walker B mutant of rhodococcal Cpa |
|
| Recombinant DNA reagent |
pET28a-His6-TEV -mKO2-RerCpa |
This paper | Plasmid for expression of mKO2-Cpa fusion protein used in MST experiments |
|
| Recombinant DNA reagent |
pETDuet-RerPrcA -PrcB∆pro |
This paper | Plasmid for expression of rhodococcal proteasome (closed-gate variant) |
|
| Recombinant DNA reagent |
pETDuet-RerPrcA∆7 -PrcB∆pro |
This paper | Plasmid for expression of rhodococcal proteasome (open-gate variant) |
|
| Recombinant DNA reagent |
pET28a-His6-TEV-RerArc | This paper | Plasmid for expression of rhodococcal Arc |
|
| Recombinant DNA reagent |
pETDuet-MtbPanB-Strep | PMID: 20203624 | ||
| Recombinant DNA reagent |
pTrc99-MtbMpa | PMID: 20203624 | ||
| Recombinant DNA reagent |
pET20-His6-Trx- TEV-MtbPup-GFPuv |
PMID: 20203624 | ||
| Recombinant DNA reagent |
pKT25 | Euromedex | ||
| Recombinant DNA reagent |
pUT18 | Euromedex | ||
| Recombinant DNA reagent |
pKT25-zip | Euromedex | ||
| Recombinant DNA reagent |
pUT18C-zip | Euromedex | ||
| Recombinant DNA reagent |
pKT25-MtbCpa | This paper | Bacterial two-hybrid plasmid: T25-Cpa fusion |
|
| Recombinant DNA reagent |
pKT25-MtbCpa∆C5 | This paper | Bacterial two-hybrid plasmid: T25-Cpa fusion where Cpa is C-terminally truncated by five residues |
|
| Recombinant DNA reagent |
pUT18-MtbPrcA- PrcB∆pro |
This paper | Bacterial two-hybrid plasmid: mycobacterial proteasome with the T18 subdomain fused to the C-terminus of PrcA |
|
| Commercial assay or kit |
Monolith NT.115 Premium Coated Capillaries |
NanoTemper Technologies |
MO-K005 | |
| Chemical compound, drug |
4-Methylumbelliferone | Sigma-Aldrich | M1381 | |
| Chemical compound, drug |
4-Methylumbelliferyl β-D-galactopyranoside |
Sigma-Aldrich | M1633 | |
| Software, algorithm |
STRING | PMID: 27924014 | ||
| Software, algorithm |
DAVID | PMID: 22543366 | ||
| Software, algorithm |
FastTree | PMID: 20224823 | ||
| Software, algorithm |
SWISS-MODEL | PMID: 28874689 | ||
| Software, algorithm |
PyMOL | Schrödinger, LLC | http://www.pymol.org |