Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | CAMK2N1 | GenBank | HGNC:HGNC:1460 | |
| Strain, strain background (Escherichia coli) | BL21(DE3) | Sigma-Aldrich | CMC0014 | Chemically competent |
| Cell line (Homo sapiens) | HEK293T | UC Berkeley cell culture facility | Authenticated using STR profiling and tested negative for mycoplasma | |
| Recombinant DNA reagent | pEGFP-C1 (plasmid) | Clontech, Mountain View, CA | Vector backbone for inserting the CaMKII genes | |
| Recombinant DNA reagent | pSNAPf (plasmid) | New England Biolabs, MA | N9183S | Vector backbone |
| Transfected construct (Homo sapiens) | pEGFP-C1-Avi-mEGFP-CaMKII | This paper | pEGFP-C1 vector was modified to have a biotinylation sequence followed by a linker at the N-terminus of mEGFP. Full-length human CaMKII-α is at the C-terminus of mEGFP. | |
| Transfected construct (Homo sapiens) | pET21a-BirA | Addgene | # 20857 | |
| Transfected construct | pcDNA3.1 | Addgene | ||
| Recombinant DNA reagent | pET 28-6xHis-precission- CaMKII Hub | This paper | pET-28 vector was modified to contain a PreScission Protease site between the N-terminal 6-histidine tag and the Human CaMKII-α hub coding sequence | |
| Recombinant DNA reagent | pSNAPf- BirA | This paper | BirA was cloned into the pSNAPf vector after removing the SNAP-tag. | |
| Peptide, recombinant protein | Peptide A (LKKFNARRKLKGAILTTMLA) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide B (AILTTMLATRNFSGGKSGGNKK) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide C (LKEFNAERKLEGAILTTMLA) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide D (LKKFNARRKLKGAGSGSGSAC) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide G (RSTVASCMHRQETVDCLKKFNA) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide F (LKKFNARRKLKGAILGSMLAC) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide H, CaMKIINTide (KRPPKLGQIGRSKRVVIEDDRIDDVLK) |
Elim Biopharm | ||
| Peptide, recombinant protein | Peptide Aphos(LKKFNARRKLKGAILpTTMLA) | Elim Biopharm | ||
| Peptide, recombinant protein | Peptide Bphos(AILpTTMLATRNFSGGKSGGNKK) | Elim Biopharm | ||
| Peptide, recombinant protein | Poly-L-lysine PEG (PLL:PEG) | SuSoS, Dübendorf, Switzerland | PLL(20)-g[3.5]- PEG(2) | Preparation of flow chambers |
| Peptide, recombinant protein | streptavidin | Sigma-Aldrich | S0677 | Functionalize the glass substrates for capturing biotinylated CaMKII |
| Peptide, recombinant protein | Calmodulin | Sigma-Aldrich | C4874 | Activation of CaMKII |
| Chemical compound, drug | PEG-Biotin | SuSoS, Dübendorf, Switzerland | PLL(20)-g[3.5]- PEG(2)/PEG(3.4)- biotin(50%) | Preparation of flow chambers |
| Chemical compound, drug | 1% protease inhibitor cocktail | Sigma | P8340 | Protease inhibitor cocktail for lysis buffer |
| Chemical compound, drug | 0.5% phosphatase inhibitor cocktail 2 and 3 | Sigma | P0044 and P5726 | Phosphatase inhibitor cocktails for lysis buffer |
| Software, algorithm | FIJI (ImageJ) | Open access software, see https://imagej.net/Fiji/Downloads | Image processing | |
| Software, algorithm | In-house python code | Open access, see, See Figure 8 and 9 Figure 8—source code 1 | Image processing | |
| Other | Sticky-Slide VI 0.4 | Ibidi | 80608 | Flow chambers |
| Other | Glass coverslips | Ibidi | 10812 | functionalized substrates |
| Software, algorithm | Mass spectrometry data acquisition | Software MassLynx 4.1 SCN 957 | Data collected on a Waters SYNAPT G2Si Mass spectrometer. Version 4.1 SCN 957 or later required to process data |
|
| Other | 500 nm opening borosilicate emitters | Generated in the lab with a tip puller |