KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-PACRG mouse monoclonal antibody (C-8) | Santa Cruz Biotechnology | Cat: sc-373851; RRID: AB_10988029 |
| Anti-His-tag rabbit polyclonal antibody | Cell Signaling Technology | Cat: 2366; RRID: AB_2115719 |
| Anti-TAMRA mouse monoclonal antibody (11H10) | ThermoFisher Scientific | Cat: MA1-042; RRID: AB_2536729 |
| Bacterial and Virus Strains | ||
| E. coli BL21(DE3) – Novagen | Sigma-Aldrich/ Merck | Cat: 69450-3 |
| E. coli DH5α | New England Biolabs | Cat: C2987I |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Restriction endonucleases: BamHI, NcoI, SalI, XhoI | New England Biolabs | Cat: BamHI R0136S, NcoI, R0193S, SalI R0138S, XhoI R0146S |
| T4 DNA ligase | New England Biolabs | Cat: M0202T |
| QuickChange II Site Directed Mutagenesis kit | Agilent | Cat: 200524 |
| p-iodo-L-phenylalanine | Sigma-Aldrich | Cat: I8757 |
| Ampicillin sodium salt | Fisher Scientific | Cat: BP176025 |
| Anhydrotetracycline | Sigma-Aldrich | Cat: 37919 |
| Chloramphenicol | Biobasic | Cat: CB0118 |
| IPTG | Biobasic | Cat: IB0168 |
| Ni-NTA agarose | Qiagen | Cat: 1000632 |
| Glutathione-sepharose 4B | Cytiva | Cat: 17-0756-01 |
| Iodoacetamide | Sigma-Aldrich | Cat: I6125 |
| Subtilisin | Sigma-Aldrich | Cat: P5380 |
| Trypsin | Sigma-Aldrich | Cat: T6567 |
| GST-3C protease | Homemade | N/A |
| His6-TEV protease | Homemade | N/A |
| JCSG plus screen | Molecular Dimensions | Cat: MD-137 |
| 50% w/v PEG 8000 | Molecular Dimensions | Cat: MD2-100-13 |
| 100% glycerol | Molecular Dimensions | Cat: MD2-100-65 |
| 1 M potassium phosphate monobasic | Molecular Dimensions | Cat: MD2-100-164 |
| Fractogel EMD SO3- (M) resin | Millipore-Sigma | Cat: 1168820010 |
| HiLyte Fluor 488 acid NHS ester | Anaspec | Cat: AS-81161 |
| Tetramethylrhodamine (TAMRA) | ThermoFisher Scientific | Cat:C1171 |
| Guanosine-5’-[(α,β)-methyleno]triphosphate (GMPCPP) | Jena Bioscience | Cat: NU-405S |
| Tubulin | Cytoskeleton | Cat: T240-A |
| PIPES | Sigma-Aldrich | Cat: 80635 |
| Paclitaxel from Taxus brevifolia, PTX; Taxol | Gold Bio | Cat: P-925-100 |
| Guanosine triphosphate (GTP) | Sigma-Aldrich | Cat: G5884 |
| 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) | ThermoFisher Scientific | Cat: PG82079 |
| N-hydroxysuccimide (NHS) | ThermoFisher Scientific | Cat: PI24500 |
| Disuccinimidyl suberate (DSS) | ThermoFisher Scientific | Cat: 21655 |
| Critical Commercial Assays | ||
| 4-20% stain-free TGX gradient gels | BioRad | Cat: 4568096 |
| Pepclean C18 spin columns | ThermoFisher Scientific | Cat: PI89873 |
| Deposited Data | ||
| Native MEIG1:PACRGΔ1–69 | This work | PDB: 6NDU |
| iodo-Y189Φ MEIG1:PACRGΔ1–69 | This work | PDB: 6NEP |
| Proteolysed MEIG1:PACRGFL | This work | PDB: 6UCC |
| Porcine tubulin dimer | (Nogales et al., 1998) | PDB: 1TUB |
| Chlamydomonas reinhardtii axonemal doublet microtubule | (Khalifa et al., 2020) | PDB: 6VE7 EMD: 20858 |
| Recombinant DNA | ||
| Human His6-MEIG1, cloned in pRSET-A | This work/Thermofisher | N/A |
| Human PACRG1–257 (FL), cloned in pGEX-6P-1 | This work/DNA Express | N/A |
| Human PACRGΔ1–69 WT, cloned in pGEX-6P-1 | This work | N/A |
| Human PACRGΔ1–69 F75X, cloned in pGEX-6P-1 | This work | N/A |
| Human PACRGΔ1–69 Y76X, cloned in pGEX-6P-1 | This work | N/A |
| Human PACRGΔ1–69 Y108X, cloned in pGEX-6P-1 | This work | N/A |
| Human PACRGΔ1–69 F121X, cloned in pGEX-6P-1 | This work | N/A |
| Human PACRGΔ1–69 Y189X, cloned in pGEX-6P-1 | This work | N/A |
| Human PACRG-WT His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG Δ1–35 His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG Δ36–69 His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-Y76F His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E77K His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E86A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E86K His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-H87A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-D88A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-K93I His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-A95T His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E99A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-I100E His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E101A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-H106A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-H132A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E136A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E136Q His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-H137A His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-R160Q His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-P195R His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-E252K His6MEIG1, cloned in pRSF-DUET | This work | N/A |
| Human PACRG-Y76F-E77K-E86KH87A His6MEIG1, cloned in pRSF-DUET (Group 1) | This work | N/A |
| Human PACRG-K154E-R160Q-R191E His6MEIG1, cloned in pRSF-DUET (Group 2) | This work | N/A |
| Human PACRG-WT His6MEIG1-H13AK78A-H81A-K82A-Y86A, cloned in pRSF-DUET (Group 3) | This work | N/A |
| Human PACRGL1–248 (FL), cloned in pET-151D-TOPO | This work/Thermofisher | N/A |
| pDB070.iodo.5 | (Bryson et al., 2017) / Addgene | Cat: 99397 |
| Software and Algorithms | ||
| XDS | (Kabsch, 2010) | xds.mpimf-heidelberg.mpg.de |
| Aimless (CCP4 package) | (Winn et al., 2011) | www.ccp4.ac.uk/html/aimless.html |
| PHENIX 1.12 including Autosol, PHASER-MR, Refine | (Liebschner et al., 2019) | www.phenix-online.org/documentation/index.html |
| ARP/wARP 8.0 | (Langer et al., 2008) | www.embl-hamburg.de/ARP |
| Coot 0.8.9.1 | (Emsley et al., 2010) | www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot |
| Fiji | (Schindelin et al., 2012) | imagej.net/Fiji |
| DataAnalysis and Biotools | Bruker | www.bruker.com |
| PyMOL 2.4.0 | Schrödinger LLC | pymol.org |
| Prism 9 | GraphPad | www.graphpad.com/scientificsoftware/prism |
| Excel 16.40 | Microsoft | www.microsoft.com/fr-ca/microsoft365/excel |
| GnomAD server | (Lek et al., 2016) | gnomad.broadinstitute.org |
| I-TASSER server | (Yang et al., 2015) | zhanglab.ccmb.med.umich.edu/ITASSER |
| Protein Atlas server | (Uhlen et al., 2015) | www.proteinatlas.org |
| Other | ||
| GSTrap 4B | Cytiva | Cat: 45001429 |
| HiLoad 16/600 Superdex 75 pg | Cytiva | Cat: 28989333 |
| Superdex 75 Increase 10/300 GL | Cytiva | Cat: 29148721 |