KEY RESOURCES TABLE
| Reagent or resource | Source | Identifier |
|---|---|---|
| Antibodies | ||
|
| ||
| Mouse monoclonal anti-alpha-Tubulin | Sigma-Aldrich | Cat# T9026; RRID: AB_477593 |
| Mouse monoclonal anti-GAPDH | Invitrogen | Cat# MA5–15738; RRID: AB_10977387 |
| Mouse monoclonal anti-Calreticulin | Abcam | Cat# ab22683; RRID: AB_447253 |
| Rabbit polyclonal anti-TAOK2 | Sigma-Aldrich | Cat# HPA010650; RRID: AB_1080204 |
| Rabbit polyclonal anti-Rtn3a | ProteinTech | Cat# 12055–2-AP; RRID: AB_2301357 |
| Mouse monoclonal anti-Acetylated alpha Tubulin | Sigma-Aldrich | Cat# T6793; RRID: AB_477585 |
| Mouse monoclonal anti-GST | Invitrogen | Cat# 13–6700; RRID: AB_2533028 |
| Mouse monoclonal anti-GM130 | BD Labs | Cat# 610822; RRID: AB_398141 |
| Rabbit polyclonal anti-TAOK2-Cterm | This paper | N/A |
| Mouse monoclonal anti-Stim1 | Santa Cruz Biotech | sc-166840; RRID: AB_2198006 |
| Mouse anti-GFP | Roche | Cat# 11814460001; RRID: AB_390913 |
| Rabbit polyclonal anti-Phospho-TAO2 (S181) | R&D Systems | Cat# PPS037; RRID: AB_2255678 |
| Mouse monoclonal anti-Tom20 | Santa Cruz Biotech | Cat# sc-17764; RRID: AB_628381 |
| Mouse monoclonal anti-EB1 | Santa Cruz Biotech | Cat# sc-47704; RRID: AB_2141631 |
| Rabbit anti-Alpha Tubulin | Cell Signaling | Cat# 2144; RRID: AB_2210548 |
|
| ||
| Bacterial strains | ||
|
| ||
| BL21 (DE3) E. coli | Invitrogen | Cat #EC0114 |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Lipofectamine 2000 reagent | Invitrogen | Cat#11668–030 |
| Purified tubulin (>99%) Porcine Brain | Cytoskeleton Inc. | Cat# T240-A |
| Halt Phosphatase Inhibitors | Thermo Fisher | Cat# 78428 |
| CP43 | Tocris | Cat#6558, CAS 850467–66-2 |
| GST-TAOK2C (1187–1235) | This paper | N/A |
|
| ||
| Experimental models: Cell lines | ||
|
| ||
| HEK293T | ATCC | Cat# CRL-3216 |
| HEK TAOK2 KO1 | This paper | N/A |
| HEK TAOK2 KO2 | This paper | N/A |
| HeLa | ATCC | Cat# CCL-2 |
|
| ||
| Oligonucleotides | ||
|
| ||
| TAOK2 KO gRNA 1: GAAATGGCAAGACATCATCA |
This paper | N/A |
| TAOK2 KO gRNA 2: ATGGCAAGACATCATCAAGG |
This paper | N/A |
| TAOK2 KO gRNA 3: TGCGGTTCTTACAGAAGCTC |
This paper | N/A |
| TAOK2 KO gRNA 4: AGTACCGGGGCTGTTACCTG |
This paper | N/A |
|
| ||
| Recombinant DNA | ||
|
| ||
| mCh-Sec61 beta | Addgene | Cat# 49155 |
| sfGFP-C1 | Addgene | Cat# 54579 |
| ER-mRFP | Addgene | Cat# 62236 |
| EGFP-Sec22b | Addgene | Cat# 101918 |
| BFP-Sec61 | Addgene | Cat# 49154 |
| Stim1-YFP | Addgene | Cat# 19754 |
| EB3-mcherry | Addgene | Cat# 55037 |
| mito-BFP | Addgene | Cat# 49151 |
| mTagBFP2-Lysosomes20 | Addgene | Cat# 55308 |
| Rab5-BFP | Addgene | Cat# 49147 |
| mEmerald Ensconsin-C-18 | Addgene | Cat# 62753 |
| sfGFP-TAOK2 | This paper | Origene CAT#: RC214297 |
| GST-TAOK2-(1187–1235) | This paper | N/A |
| BFP-STIM1 | This paper | From Cat# 19754 |
|
| ||
| Software and algorithms | ||
|
| ||
| GraphPad software | Prism 9.0 | https://www.graphpad.com/scientific-software/prism/ |
| TMHMM2.0 | Krogh et al., 2001 | https://services.healthtech.dtu.dk/service.php?TMHMM-2.0 |
| Heliquest | Gautier et al., 2008 | https://heliquest.ipmc.cnrs.fr/ |
| AMPHIPASEEK | Combet et al., 2000 | https://npsa-prabi.ibcp.fr/NPSA/npsa_amphipaseek.html |
| AlphaFold2.0 | Jumper et al., 2021 | https://alphafold.ebi.ac.uk/ |