IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ CASK Functions as a Mg2+-independent neurexin kinase. CASK Functions as a Mg2+-independent neurexin kinase. Cell (0092-8674) 2008 imex curation Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu 08-Jul-2008: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu Accepted 2008-JUL-13 by DAVET. 08-Jul-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 4 imex curation CASK Functions as a Mg2+-independent neurexin kinase. CASK Functions as a Mg2+-independent neurexin kinase. Cell (0092-8674) 2008 imex curation Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu 08-Jul-2008: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting predetermined predetermined participant predetermined Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. The nucleotide was cross-linked with protein using UV-Stratalinker. konark.mukherjee@utsouthwestern.edu Accepted 2008-JUL-13 by DAVET. 08-Jul-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 4 imex curation CASK Functions as a Mg2+-independent neurexin kinase. CASK Functions as a Mg2+-independent neurexin kinase. Cell (0092-8674) 2008 imex curation Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu 08-Jul-2008: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. Details of origin of Nrxn1 supplied by author konark.mukherjee@utsouthwestern.edu Accepted 2008-JUL-13 by DAVET. 08-Jul-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 4 imex curation CASK Functions as a Mg2+-independent neurexin kinase. CASK Functions as a Mg2+-independent neurexin kinase. Cell (0092-8674) 2008 imex curation Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu 08-Jul-2008: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. Details of origin of Nrxn1 supplied by author konark.mukherjee@utsouthwestern.edu Accepted 2008-JUL-13 by DAVET. 08-Jul-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 4 imex curation CASK Functions as a Mg2+-independent neurexin kinase. CASK Functions as a Mg2+-independent neurexin kinase. Cell (0092-8674) 2008 imex curation Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu 08-Jul-2008: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu Accepted 2008-JUL-13 by DAVET. 08-Jul-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 4 imex curation CASK Functions as a Mg2+-independent neurexin kinase. CASK Functions as a Mg2+-independent neurexin kinase. Cell (0092-8674) 2008 imex curation Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu 08-Jul-2008: Contacted by JYOTI. davet Only protein-protein interactions 4 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. Details of origin of Nrxn1 supplied by author konark.mukherjee@utsouthwestern.edu Accepted 2008-JUL-13 by DAVET. 08-Jul-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 4 imex curation tnp-atp 2',3'-O-(2,4,6- trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ChEBI entry requested ChEBI entry requested nrx1b_rat Neurexin-1-beta Nrxn1 Neurexin I-beta protein protein rat Rattus norvegicus (Rat) Rat MYQRMLRCGAELGSPGGGSSGGAGGRLALLWIVPLTLSGLLGVAWGASSLGAHHIHHFHGSSKHHSVPIAIYRSPASLRGGHAGTTYIFSKGGGQITYKWPPNDRPSTRADRLAIGFSTVQKEAVLVRVDSSSGLGDYLELHIHQGKIGVKFNVGTDDIAIEESNAIINDGKYHVVRFTRSGGNATLQVDSWPVIERYPAGNNDNERLAIARQRIPYRLGRVVDEWLLDKGRQLTIFNSQATIIIGGKEQGQPFQGQLSGLYYNGLKVLNMAAENDANIAIVGNVRLVGEVPSSMTTESTATAMQSEMSTSIMETTTTLATSTARRGKPPTKEPISQTTDDILVASAECPSDDEDIDPCEPSSGGLANPTRVGGREPYPGSAEVIRESSSTTGMVVGIVAAAALCILILLYAMYKYRNRDEGSYHVDESRNYISNSAQSNGAVVKEKQPSSAKSANKNKKNKDKEYYV DIP protein Q63373 original sequence version: 92 B24A3CEFF5A2394E adenosine 3'-monophosphate adenosine 3'-monophosphate (3'-AMP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cskp_human Peripheral plasma membrane protein CASK CASK LIN2 Calcium/calmodulin-dependent serine protein kinase Protein lin-2 homolog protein protein human Homo sapiens Human MADDDVLFEDVYELCEVIGKGPFSVVRRCINRETGQQFAVKIVDVAKFTSSPGLSTEDLKREASICHMLKHPHIVELLETYSSDGMLYMVFEFMDGADLCFEIVKRADAGFVYSEAVASHYMRQILEALRYCHDNNIIHRDVKPHCVLLASKENSAPVKLGGFGVAIQLGESGLVAGGRVGTPHFMAPEVVKREPYGKPVDVWGCGVILFILLSGCLPFYGTKERLFEGIIKGKYKMNPRQWSHISESAKDLVRRMLMLDPAERITVYEALNHPWLKERDRYAYKIHLPETVEQLRKFNARRKLKGAVLAAVSSHKFNSFYGDPPEELPDFSEDPTSSGLLAAERAVSQVLDSLEEIHALTDCSEKDLDFLHSVFQDQHLHTLLDLYDKINTKSSPQIRNPPSDAVQRAKEVLEEISCYPENNDAKELKRILTQPHFMALLQTHDVVAHEVYSDEALRVTPPPTSPYLNGDSPESANGDMDMENVTRVRLVQFQKNTDEPMGITLKMNELNHCIVARIMHGGMIHRQGTLHVGDEIREINGISVANQTVEQLQKMLREMRGSITFKIVPSYRTQSSSCERDSPSTSRQSPANGHSSTNNSVSDLPSTTQPKGRQIYVRAQFEYDPAKDDLIPCKEAGIRFRVGDIIQIISKDDHNWWQGKLENSKNGTAGLIPSPELQEWRVACIAMEKTKQEQQASCTWFGKKKKQYKDKYLAKHNAVFDQLDLVTYEEVVKLPAFKRKTLVLLGAHGVGRRHIKNTLITKHPDRFAYPIPHTTRPPKKDEENGKNYYFVSHDQMMQDISNNEYLEYGSHEDAMYGTKLETIRKIHEQGLIAILDVEPQALKVLRTAEFAPFVVFIAAPTITPGLNEDESLQRLQKESDILQRTYAHYFDLTIINNEIDETIRHLEEAVELVCTAPQWVPVSWVY 6C02008CE52728BA atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) amp-pnp AMP-PNP small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cask-amp 1 9 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 1.85A working 20.2%, free 26.7% Fig. 1, Table S1, Supp. Fig. 3 Ribbon diagrams depicting the overall fold of the CASK CaM-kinase domain in a complex with 3'-AMP cask-amppnp-1 1 12 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 2.19A working 20.2%, free 25.7% Fig. 1B, 2B, Table S1, Supp. Fig. 2 Stereo plots of the nucleotide-binding pockets of the CASK (B, in complex with AMPPNP; triclinic form) cask-tnp-atp 5 10 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Fig. 3A-D We first tested whether the CASK CaM-kinase domain interacts with the ATP analog TNP-ATP that becomes fluorescent when inserted into the hydrophobic ATP-binding pocket ATP titrations revealed that a 500-fold excess of ATP reduced the TNP-ATP fluorescence by 50% (KiATP = 0.365 mM), consistent with competitive binding of ATP and TNP-ATP to a single site that exhibits a much higher affinity for TNP-ATP than for ATP. Divalent cations could also inhibit this interaction, but the authors speculate that this was by complexing with ATP rather than by binding to the enzyme. EDTA stimulates the interaction, presumably by chelating divalent cations. cask-1 4 10 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain O14936:p.[Ser24Asp;Val26Leu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain V L Wed May 02 15:25:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser24asp,val26leu' Tue Jun 30 17:48:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser24Asp;Val26Leu]' ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation true KmATP of =0.563 mM, Vmax = 4.9 nmol/umol enzyme/min KiMg2+ = 61 uM Figs. 4A-D, Supp. Fig. S11A Phosphorylation of Ser-151 and Ser-155 identified by mass spectrometry Mg2+ and other divalent cations inhibited but did not abolish the autophosphorylation of CASK, presumably by reducing its ATP affinity cask-nrxn1-1 4 10 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain O14936:p.[Ser24Asp;Val26Leu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain V L certain certain sequence position certain certain certain sequence position certain S D Fri Jul 22 10:50:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'val26leu,ser24asp' Wed May 02 14:32:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser24asp,val26leu' Tue Jun 30 19:24:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser24Asp;Val26Leu]' ecobb Escherichia coli (strain B / BL21) 8 enzyme target enzyme target substrate substrate neutral component neutral component c-terminal tail sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation KmATP of neurexin-1 phosphorylation by CASK (=748.7 uM, Vmax = 159.4 nmol/umol enzyme/min Fig. 5, Supp. Figs. 10, 11B Phosphorylation of Nrxn1 identified by mass spectrometry cask-atp-1 2 11 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Supp. Fig. 6A ATP binding to the CASK CaM-kinase domain ATP binding is inhibited by Mg2+ but not by Na+. cask-nrxn1-2 6 10 unspecified role unspecified role neutral component neutral component region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component c-terminal tail sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain cherry tag region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Supp. Fig. S8A coexpression of a CASK-EGFP fusion protein with neurexin-1β in HEK293 cells recruits CASK from the cytosol to the plasma membrane Q63373; O14936 transfection cask-nrxn1-3 3 8 unspecified role unspecified role bait bait c-terminal tail sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 10 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Supp. Fig. 9A GST-neurexin C-tail (GST-Nx-Ct) or GST alone were immobilized on glutathione beads, and incubated with full-length CASK, Binding does not appear to be dependent on adenine-nucleotides