Table 3. Key reagents and resources.
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rabbit polyclonal anti-CSPα (used at 1:5,000) | Chemicon | Cat# AB1576; RRID: AB_90794 |
Rabbit polyclonal anti-endophilin 1 (used at 1:1,000) | Synaptic Systems | Cat# 159 002; RRID: AB_887757 |
Mouse monoclonal anti-SNAP-25 (used at 1:5,000) | Covance | Cat# SMI-81; RRID: AB_2315336 |
Mouse monoclonal anti-β-Actin (used at 1:1,000) | GeneTex | Cat# GTX629630; RRID: AB_2728646 |
Mouse monoclonal anti-dynamin (used at 1:2,000) | BD Biosciences | Cat# 610246; RRID: AB_397641 |
Rabbit polyclonal anti-NrCAM (used at 1:10,000) | Abcam | Cat# ab24344; RRID: AB_448024 |
Rabbit polyclonal anti-ATP1B2 (used at 1:200) | Thermo Fisher Scientific | Cat# PA5-11852; AB_2857261 |
Rabbit polyclonal anti-GluA1 (used at 1:1,000) | Millipore | Cat# AB1504; RRID: AB_2113602 |
Rat anti-Syncam2 (used at 1:2,000 for ICC) | Gift from the laboratory of Dr. Thomas Biederer | N/A–Custom Antibody |
Chicken polyclonal anti-MAP2 (used at 1:500 for ICC) | Millipore | Cat# AB5543; RRID: AB_571049 |
Mouse monoclonal anti-myelin basic protein (MBP) (used at 1:750) | Abcam | Cat# ab62631; RRID: AB_956157 |
Rabbit polyclonal anti-synaptogyrin 3 (used at 1:1,000) | Synaptic Systems | Cat# 103 3030; RRID: AB_2619753 |
Mouse monoclonal anti-synaptophysin 1 (used at 1:10,000 for WB, 1:500 for ICC) | Synaptic Systems | Cat# 101 011; RRID: AB_887822 |
Rabbit monoclonal anti-α-Synuclein (used at 1:1,000) | Cell Signaling Technology | Cat# 4179; RRID: AB_1904156 |
Rabbit polyclonal anti-PPT1 (used at 1:200 for ICC) | GeneTex | Cat #: GTX110677; RRID: AB_1951370 |
Rabbit polyclonal anti-PPT1 (used at 1:50 for WB) | This paper; Thermo Fisher Scientific Life Sciences Solutions | Custom Antibody; Project VH2380 |
IRDye 680RD goat anti-mouse IgG secondary | LI-COR Biosciences | Cat# 926–68070; RRID: AB_10956588 |
IRDye 680RD goat anti-rabbit IgG secondary | LI-COR Biosciences | Cat# 926–68071: RRID: AB_10956166 |
IRDye 800CW goat anti-rabbit IgG secondary | LI-COR Biosciences | Cat# 926–32211; RRID: AB_621843 |
IRDye 800CW donkey anti-mouse IgG secondary | LI-COR Biosciences | Cat# 926–32212; RRID: AB_621847 |
Goat anti-rat IgG (H+L) cross-adsorbed, Alexa Fluor 488 | Invitrogen | Cat# A11006; RRID: AB_141373 |
Goat polyclonal anti-chicken IgY (H+L) cross-adsorbed, Alexa Fluor 633 secondary | Invitrogen | Cat# A21103; RRID: AB_2535756 |
Chemicals, peptides, and recombinant proteins | ||
TCEP | Thermo Fisher Scientific | Cat# 20490 |
HA | Millipore Sigma | Cat# 467804 |
NEM | Thermo Fisher Scientific | Cat# 23030 |
Thiopropyl sepharose 4B | Millipore Sigma | Cat# T8512 |
dNEM | Cambridge Isotope Laboratories | Cat# DLM-6711-10 |
QuikChange II XL Site-Directed Mutagenesis Kit | Agilent | Cat# 200521 |
4-methylumbelliferyl-6-thio-Palmitate-β-D-glucopyranoside (MU-6S-Palm-βGlc) | Carbosynth | Cat# EM06650 |
β-glucosidase from almonds | Sigma | Cat# 49290 |
Deposited data | ||
Analyzed mass spectrometry data | This paper | Tables |
Raw data | ProteomeXchange Consortium | PXD017270 |
Experimental models: Cell lines | ||
HEK 293T cells | American Type Culture Collection | Cat# CRL-3216 |
Experimental models: Organisms/strains | ||
Mus musculus—B6;129-Ppt1tm1Hof/J | The Jackson Labs | JAX: 004313 |
Xenopus laevis | Nasco | LM00531 |
Recombinant DNA | ||
LCV-hPPT1 | This paper | N/A |
LCV-FUGW | This paper | N/A |
pGEMHE-GluA1.WT | This paper | N/A |
pGEMHE-GluA1.C323A | This paper | N/A |
pGEMHE-TARPg8 | This paper | N/A |
Software and algorithms | ||
CSS Palm | Ren and colleagues (2008) | http://csspalm.biocuckoo.org |
Swiss Palm | Blanc and colleagues (2015) | https://swisspalm.org |
IPA | Qiagen | https://www.qiagenbioinformatics.com/products/ingenuity-pathway-analysis |
GraphPad Prism 7 | GraphPad | https://www.graphpad.com/scientific-software/prism |
Progenesis QI Proteomics software V4.0 | Nonlinear Dynamics | http://www.nonlinear.com/progenesis/qi-for-proteomics/download |
Mascot search algorithm | Matrix Science | http://www.matrixscience.com/search_form_select.html |
WebLogo | Crooks and colleagues (2004) | https://weblogo.berkeley.edu |
SWISS-MODEL | Bienert and colleagues (2017) | https://swissmodel.expasy.org |
Sourcing and catalog information for materials and resources used to conduct this study.
dNEM, d5-N-ethylmaleimide; HA, hydroxylamine; IPA, Ingenuity Pathway Analysis; NEM, N-ethylmaleimide; TCEP, tris(2-carboxyethyl)phosphine.