Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | CDK2 (rabbit polyclonal) |
Santa Cruz | sc-163 | WB (1:1000) |
| Antibody | Cyclin A (mouse monoclonal) |
Santa Cruz | sc-271682 | WB (1:1000) |
| Antibody | Flag (mouse monoclonal) |
Sigma | A8592-.2MG | WB (1:2500, 1:10,000) |
| Antibody | Flag (mouse monoclonal) |
GenScript | A00187 | IP (1 µg/mL) |
| Antibody | GFP (rabbit monoclonal) |
CST | 2956S | WB (1:1000) |
| Antibody | GST (mouse monoclonal) |
Santa Cruz | sc-138 | WB (1:1000) |
| Antibody | PP2A Aβ subunit (goat polyclonal) |
Santa Cruz | sc-6113 | WB (1:2000) |
| Antibody | PP2A B subunit (100C1) (rabbit monoclonal) |
CST | 2290S | WB (1:2000) |
| Antibody | PP2A C subunit (1D6) (mouse monoclonal) | Millipore | 05-421 | WB (1:5000) |
| Antibody | Phospho-CDK Substrate Motif (rabbit monoclonal) |
CST | 9477S | WB (1:1000) |
| Antibody | ECL Rabbit IgG, HRP-linked whole Ab (from donkey) | GE Healthcare | NA934V | WB/secondary antibody (1:10,000) |
| Antibody | HA (12CA5) (mouse monoclonal) |
Roche/Sigma | 11583816001 | WB (1:500) |
| Antibody | ECL Mouse IgG, HRP-linked whole Ab (from donkey) | GE Healthcare | NA931V | WB/secondary antibody (1:10,000) |
| Antibody | Mouse anti-goat IgG-HRP | Santa Cruz | sc-2354 | WB/secondary antibody (1:10,000) |
| Antibody | Monoclonal ANTI-FLAG M2 antibody produced in mouse, ANTI-FLAG M2 Affinity Agarose Gel | Sigma | A2220 | IP (10 μL) |
| Antibody | Anti-c-Myc Agarose Affinity Gel antibody produced in rabbit (polyclonal) | Sigma | A7470 | IP (10 μL) |
| Antibody | GFP-Trap agarose beads | Chromotek | gta-10 | IP (5 μL) |
| Peptide, recombinant protein | Synthetic peptides for competition assays | Biomatik | Custom | Sequence variant described in this paper |
| Peptide, recombinant protein | DYKDDDDK peptide | GenScript | RP10586 | |
| Peptide, recombinant protein | Recombinant cyclin A/CDK2 | Thermo Fisher | PV3267 | |
| Peptide, recombinant protein | Purified recombinant B55α/PP2A holoenzyme | Zhao et al., 2019 | Trimeric Flag-B55α/PP2A holoenzyme purified from 293T cells. | |
| Strain, strain background (Escherichia coli) | BL21-Gold (DE3) cells | Agilent | 230132 | To generate GST-Fusion proteins |
| Cell line (Homo sapiens) | 293T cells | ATCC | CRL-3216 | Transient transfections and source of cell lysates |
| Cell line (H. sapiens) | U-2 OS cells | ATCC | HTB-96 | Transient transfections |
| Cell line (H. sapiens) | Expi293F cells | Thermo Fisher | A14528 | Purification of recombinant B55α |
| Transfected construct (human) | Flag-B55α 293T cells | Zhao et al., 2019 | 293T cells stably transfected with pCPP-Flag-B55α and selected with puromycin | |
| Commercial assay or kit | (AminoLink Plus Immobilization Kit) | Thermo Fisher Scientific | 44894 | |
| Commercial assay or kit | Ser/Thr phosphatase assay kit | EMD Millipore | 17-127 | |
| Commercial assay or kit | QuikChange II | Agilent | 200521 | |
| Recombinant DNA reagent | pBOS GFP-H2B plasmid | Kanda et al., 1998 | ||
| Recombinant DNA reagent | pcDNA3.4-K-GFP-RP1B | This paper | His6-green fluorescent protein-tag, a TEV cleavage | |
| Recombinant DNA reagent | pTHMT | Peti and Page, Protein Expr. Purif. 51, 1–10 (2007) | N-terminal His6-Maltose Binding Protein (MBP)-tag, a TEV cleavage | |
| Recombinant DNA reagent | pCPP-Flag-B55α | Zhao et al., 2019 | ||
| Recombinant DNA reagent | pMSCV-puro-Myc-B55α | Jayadeva et al., 2010 | ||
| Recombinant DNA reagent | pMSCV-puro-Myc-B55α variants | This paper | Generated by site-directed mutagenesis (primer sequences provided in Appendix table) | |
| Recombinant DNA reagent | pCDNA5/FRT/TO-GFP-p107-R1 | This paper | Inserting wild-type or mutant p107-R1 gBlock Gene Fragments (IDT) into the pCDNA5/FRT/TO-GFP vector via the BamHI/Not sites | |
| Recombinant DNA reagent | pGEX-2T-p107 spacer | Jayadeva et al., 2010 | ||
| Recombinant DNA reagent | pGEX-2T-p107 spacer variants | This paper | Generated by site-directed mutagenesis (primer sequences provided in Appendix table) | |
| Recombinant DNA reagent | pCDNA5/FRT/TO-GFP-p107-R1 | This paper | Inserting a gblock in pCDNA5/FRT/TO-GFP containing 4 copies of R1 | |
| Recombinant DNA reagent | pCDNA5/FRT/TO-GFP-p107-R1-H/AxR/AV/AxxV/A | This paper | Inserting a gblock in pCDNA5/FRT/TO-GFP containing four mutant copies of R1. (gblock sequences provided in Appendix table) | |
| Recombinant DNA reagent | pSG5-puro-Flag-p107 | Kurimchak et al., 2013 | ||
| Recombinant DNA reagent | pCMV-Flag-p107 | Voorhoeve et al., 1999 | ||
| Recombinant DNA reagent | pCMV-HA-E2F4 | Ginsberg et al., 1994 | ||
| Recombinant DNA reagent | pRC-cyclin E | Addgene | #8963 | |
| Software, algorithm | ConSurf | ConSurf Ashkenazy et al., 2010 |
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| Software, algorithm | ImageJ software | |||
| Software, algorithm | FlowJo v10 software | BD Biosciences | v10.8 | |
| Software, algorithm | Clustal Omega | Sievers et al., 2011 |