Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Guinea pig polyclonal Shank3 | Synaptic Systems | 162304 | for ICC (1:500) |
| Antibody | Rabbit polyclonal Homer-1,2,3 | Synaptic Systems | 160103 | for ICC (1:500) |
| Antibody | Chicken polyclonal Bassoon | Synaptic Systems | 141016 | for ICC (1:500) |
| Antibody | Mouse monoclonal MAP2-AF488 | Merck Millipore | MAB3418X | for ICC (1:500) |
| Antibody | Mouse monoclonal GFP | Covance | MMS-118P-500 | for Western Blot (1:3000) |
| Antibody | Rat monoclonal RFP | Chromotek | 5F8 | for Western Blot (1:1000) |
| Antibody | Rabbit polyclonal IgG anti-chicken IgY (H + L)-Cy3 | Dianova | 303-165-003 | for ICC (1:500) |
| Antibody | Goat polyclonal anti-rabbit IgG (H + L)-AF568 | Life Technologies | A-11036 | for ICC (1:500) |
| Antibody | Goat polyclonal anti-guinea pig IgG-STAR 635P | Abberior | ST635P-1006 | for ICC (1:500) |
| Antibody | Goat polyclonal anti-rat IgG (H + L)-HRP |
Jackson ImmunoResearch | 112-035-062; RRID:AB_
2338133 |
for Western Blot (1:2500) |
| Antibody | Goat polyclonal anti-mouse IgG (H + L)-HRP |
Jackson ImmunoResearch | 115-035-003; RRID:AB_
10015289 |
for Western Blot (1:2500) |
| Strain, strain background (Escherichia coli) | XL10 gold | Stratagene | 200314 | for plasmid DNA production; stock amplified in the lab |
| Strain, strain background (Escherichia coli) | BL21(DE3) | ThermoFisher Scientific | K30001 | for protein purification; stock amplified in the lab |
| Strain, strain background (Escherichia coli) | Stbl2 | Invitrogen | 10268–019 | obtained from Matthias Kneussel; stock amplified in the lab |
| Cell line (Homo- sapiens) | Embryonic kidney (HEK 293T) | ATCC | CRL-3216; RRID:CVCL_0063 | for co-IP |
| Transfected construct (rat) | pET-His6-SUMO-SHANK3(1-676) WT | This paper | for bacterial protein production; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pET-His6-SUMO-SHANK3(1-676) R12C | This paper | for bacterial protein production; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pET-His6-SUMO-SHANK3(1-676) L68P | This paper | for bacterial protein production; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pHAGE2-EF1a-EGFP-SHANK3FL WT | This paper | for transfection of primary hippocampal neurons; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pHAGE2-EF1a-EGFP-SHANK3FL R12C | This paper | for transfection of primary hippocampal neurons; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pHAGE2-EF1a-EGFP-SHANK3FL L68P | This paper | for transfection of primary hippocampal neurons; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pmRFP-N3-SHANK3FL WT | doi:10.1038/ncb3487 | for transfection of HEK293 cells and co-IP | |
| Transfected construct (rat) | pmRFP-N3-SHANK3FL R12C | doi:10.1038/ncb3487 | for transfection of HEK293 cells and co-IP | |
| Transfected construct (rat) | pmRFP-N3-SHANK3FL L68P | doi:10.1038/ncb3487 | for transfection of HEK293 cells and co-IP | |
| Transfected construct (rat) | pEGFP-C1-Homer1b | This paper | for transfection of HEK293 cells and co-IP; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (rat) | pEGFP-C1-SAPAP1 | This paper (cDNA coding for rat SAPAP1 obtained from Stefan Kindler) | for transfection of HEK293 cells and co-IP; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (human) | pEGFP-C1-cortactin | This paper | for transfection of HEK293 cells and co-IP; available from Hans-Jürgen Kreienkamp | |
| Transfected construct (mouse) | mCherry-cortactin-C-12 | doi:10.1016/j.neuron.2018.01.046 | Addgene plasmid #55020 | for transfection of primary hippocampal neurons |
| Transfected construct (rat) | pFUGW-f(syn)Homer1-tdTomato-WPRE | A gift from Christian Rosenmund | BL-1034 | for transfection of primary hippocampal neurons |
| Transfected construct | pAAV-Syn-mRuby2 | doi:10.15252/embj.2018101183 | for transfection of primary hippocampal neurons | |
| Transfected construct | pMAX-GFP | Lonza Bioscience | V4XP-3012 | for transfection of primary hippocampal neurons |
| Transfected construct (human) | pETM11-T7-His6-SenP2 | EMBL Heidelberg | for recombinant protein tag removal | |
| Peptide, recombinant protein | His6-SUMO-SHANK3(1-676) WT | This paper | purified from BL21(DE3) using pET-His6-SUMO-SHANK3(1-676) WT | |
| Peptide, recombinant protein | His6-SUMO-SHANK3(1-676) R12C | This paper | purified from BL21(DE3) using pET-His6-SUMO-SHANK3(1-676) R12C | |
| Peptide, recombinant protein | His6-SUMO-SHANK3(1-676) L68P | This paper | purified from BL21(DE3) using pET-His6-SUMO-SHANK3(1-676) L68P | |
| Peptide, recombinant protein | SHANK3(1-676) WT | This paper | purified from BL21(DE3), see protein purification methods | |
| Peptide, recombinant protein | SHANK3(1-676) R12C | This paper | purified from BL21(DE3), see protein purification methods | |
| Peptide, recombinant protein | SHANK3(1-676) L68P | This paper | purified from BL21(DE3), see protein purification methods | |
| Peptide, recombinant protein | His6-SenP2 | This paper | purified from BL21(DE3); for recombinant protein tag removal; see protein purification methods | |
| Chemical compound, drug | Profinity IMAC resin, Ni2+-charged | Bio-Rad | 1560135 | for protein purification |
| Chemical compound, drug | RFP-trap agarose | Chromotek | rta-20 | for co-IP |
| Chemical compound, drug | (+)-Bicuculline | Tocris Bioscience | 0130 | for treatment of hippocampal primary neurons |
| Chemical compound, drug | Tetrodotoxin | Tocris Bioscience | 1078 | for treatment of hippocampal primary neurons |
| Chemical compound, drug | D-APV | Tocris Bioscience | 0106 | for treatment of hippocampal primary neurons |
| Chemical compound, drug | Lipofectamine 2000 | ThermoFisher Scientific | 11668019 | for transfection of primary hippocampal neurons |
| Chemical compound, drug | TurboFect transfection reagent | ThermoFisher Scientific | R0532 | for transfection of HEK293 cells |
| Commercial assay or kit | BCA protein assay | ThermoFisher Scientific | 23250 | for determination of protein concentration |
| Commercial assay or kit | Plasmid midi kit | Qiagen | 12145 | for purification of plasmid DNA |
| Commercial assay or kit | WesternBright chemiluminescence substrate | Biozym | 541003 | for western blot detection |
| Software, algorithm | VMD 1.9.3 | doi:10.1016/0263-7855(96)00018-5 | for analysis and visualization of molecular graphics | |
| Software, algorithm | Origin 2019b | OriginLab | for graphing and data analysis | |
| Software, algorithm | PyMOL 2.3.4 | Schrödinger, Inc. | for molecular visualization | |
| Software, algorithm | ImageJ/Fiji | NIH | for image processing and analysis | |
| Software, algorithm | Adobe Illustrator 2021 | Adobe Inc. | for graphing and visualization | |
| Software, algorithm | ATSAS 3.0.1 | doi:10.1107/S1600576720013412 | for analysis of SAXS data | |
| Software, algorithm | SnapGene | GSL Biotech LLC | for plasmid cloning | |
| Software, algorithm | Image Lab | Bio-Rad | for processing and analysis of western blot images | |
| Software, algorithm | POV-Ray 3.7 | Other | for ray tracing of molecular graphics | |
| Software, algorithm | MATLAB | MathWorks | for SHANK3 intensity distribution analysis (Figure 5E) |