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/ Regulation of MBK-2/DYRK by CDK-1 and the pseudophosphatases EGG-4 and EGG-5 during the oocyte-to-embryo transition. Regulation of MBK-2/DYRK by CDK-1 and the pseudophosphatases EGG-4 and EGG-5 during the oocyte-to-embryo transition. Cell (0092-8674) 2009 imex curation Cheng KC., Klancer R., Singson A., Seydoux G. Apoptosis - Interactions involving proteins with a function related to apoptosis Details of CDK1 taken from manufacturer's website. This included the presence of the CDK1 activators Cyclin B1 and B2 in the reaction mix. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 6 in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Cheng KC., Klancer R., Singson A., Seydoux G. Apoptosis - Interactions involving proteins with a function related to apoptosis Details of CDK1 taken from manufacturer's website. This included the presence of the CDK1 activators Cyclin B1 and B2 in the reaction mix. gseydoux@jhmi.edu Accepted 2010-MAR-9 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 6 imex curation Regulation of MBK-2/DYRK by CDK-1 and the pseudophosphatases EGG-4 and EGG-5 during the oocyte-to-embryo transition. Regulation of MBK-2/DYRK by CDK-1 and the pseudophosphatases EGG-4 and EGG-5 during the oocyte-to-embryo transition. Cell (0092-8674) 2009 imex curation Cheng KC., Klancer R., Singson A., Seydoux G. Apoptosis - Interactions involving proteins with a function related to apoptosis Details of CDK1 taken from manufacturer's website. This included the presence of the CDK1 activators Cyclin B1 and B2 in the reaction mix. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 6 in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Cheng KC., Klancer R., Singson A., Seydoux G. Apoptosis - Interactions involving proteins with a function related to apoptosis Details of CDK1 taken from manufacturer's website. This included the presence of the CDK1 activators Cyclin B1 and B2 in the reaction mix. gseydoux@jhmi.edu Accepted 2010-MAR-9 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 6 imex curation ccnb2_human G2/mitotic-specific cyclin-B2 CCNB2 protein protein human Homo sapiens Human MALLRRPTVSSDLENIDTGVNSKVKSHVTIRRTVLEEIGNRVTTRAAQVAKKAQNTKVPVQPTKTTNVNKQLKPTASVKPVQMEKLAPKGPSPTPEDVSMKEENLCQAFSDALLCKIEDIDNEDWENPQLCSDYVKDIYQYLRQLEVLQSINPHFLDGRDINGRMRAILVDWLVQVHSKFRLLQETLYMCVGIMDRFLQVQPVSRKKLQLVGITALLLASKYEEMFSPNIEDFVYITDNAYTSSQIREMETLILKELKFELGRPLPLHFLRRASKAGEVDVEQHTLAKYLMELTLIDYDMVHYHPSKVAAAASCLSQKVLGQGKWNLKQQYYTGYTENEVLEVMQHMAKNVVKVNENLTKFIAIKNKYASSKLLKISMIPQLNSKAVKDLASPLIGRS DIP protein O95067 original sequence version: 118 874466E1DD68A4C4 egg3_caeel Protein tyrosine phosphatase-like protein egg-3 egg-3 Egg sterile protein 3 F44F4.2 protein protein caeel Caenorhabditis elegans MRTSDSHLPLSNLARSDSIEFKDAVINEKWVHSANRRKGHLTPKAPEKKSGWFGGQKSEEELLMERVEQHELEELQTFIAQKLFRVDGIICDEETRILLVEKLMNAKEYPTINHDELAHRYGSSMAGWLRDRLVPSMSDCSSVLQRAAAEFYQNKMSDPLCNWGQLNPEHVSMVAARIAKFSEEMSSKVKWSLLVEPGKFSCHLTEFVQEFNRLDRMFVSNELSDEESLQTAFNANYLTKARSKMVPCAEFSRVKLNDGLGRLDDRNELRNGMFSDEHEFLQEEGYTAKSTYGTTDFIHANYVKGGPLLNTFICAQAPLKNTQEDFWRMVFQEKCQFIVMLNSAVDSSTLGPLDSANRNHCPYYWPRAENESLRFGSFHITCMKVDSKADPLFTITKLKVQKVGGNLLDAEFDEELFLEHWQWDWQYLGDVHWPFRVLRKARQLSTPTIVQCIDGCSKSGTLVSIETALMHFIRGSPITKSLILQSCVFVRLQRRLSVSSVLLYLFIYRVILRWIEPYVNKWYHRAALGLRFKSIGFIQKYNAMIQEFSRITPAY 587DA01F0CC92E94 cdk1_human Cyclin-dependent kinase 1 p34 protein kinase Cell division protein kinase 1 CDK1 CDC2 Cell division control protein 2 homolog CDC28A CDKN1 P34CDC2 protein protein human Homo sapiens Human MEDYTKIEKIGEGTYGVVYKGRHKTTGQVVAMKKIRLESEEEGVPSTAIREISLLKELRHPNIVSLQDVLMQDSRLYLIFEFLSMDLKKYLDSIPPGQYMDSSLVKSYLYQILQGIVFCHSRRVLHRDLKPQNLLIDDKGTIKLADFGLARAFGIPIRVYTHEVVTLWYRSPEVLLGSARYSTPVDIWSIGTIFAELATKKPLFHGDSEIDQLFRIFRALGTPNNEVWPEVESLQDYKNTFPKWKPGSLASHVKNLDENGLDLLSKMLIYDPAKRISGKMALNHPYFNDLDNQIKKM DIP protein P06493 original sequence version: 185 DIP protein P06493 original sequence version: 162 DIP protein P06493 original sequence version: 184 DIP protein P06493 original sequence version: 97 DIP protein P06493 original sequence version: 192 942D79448EFE490A egg5_caeel Inactive protein-tyrosine phosphatase egg-5 egg-5 Egg sterile protein 5 Protein-tyrosine phosphatase-like protein egg-5 R12E2.10 protein protein caeel Caenorhabditis elegans MALNSEVMFREQINAMRSQAGRKRATSLQSFCSGNTDDSSADSTDNMDMMVDYPQQKGVSCMRARFNSESTLSKSFRKKVKKLAQKDRRSKERLNGNSEEDAIEVPRGAPSTYAAPSKLRKSKALDCLVSEKPKDEGRSEDSGHGADIEMAKGHFNDVRMKVFAARTAMQVEPALVMKTRKALEMKNAVLENHQSPGAFSLHAAYKIAASAESRVGSITPCNKKVTKEAMANLIRSSYDDTEITQELLFSSKFDSKWKGRYTDIYMRRDENGKKPKRPVNGQGWVMPLKSICEKFGINSTFFTNHRIDLKSARDQVLLMRLLSHDQTSTWISDIHPEAVKNETLAEYLLRELDASTMQKRVQAFKANVLADRDRVRVAGQFYNNIRIGKRMFGAARKAKYLSTIIGGMERRFEILENSVNHIPFTHSASDNNQEKCRNPRVHCRDSTRIALQFPRGQYLGDFIHANRISGKPLFNEFIMTQAPMKNTVDDFWRMVWQEEVPYIVMLTSRKEPERCEYYWPKSPSDPAVTVPGGLRIENFGVYQAPDPLFRVTHLRLIGPDREERHVEHWQGDVNNSSNMYSPLNILRLLRNASKPVVIHDHLGVSRAACLVAAEIAICSLLRGPTYKYPVQRAVQFLRQRRPFSIETPMQYIFVHRLVAFFFRDVIGSAKELDVDYERWLQERSERMFLDDLAAPIPGYRLLSPRADPDIVRMVGRPERPNYRREAPDCVGEMPNKVAAVDGILSPAKSVFEF DB71183AB91FD14F ktna1_caeel Meiotic spindle formation protein mei-1 mei-1 Katanin p60 ATPase-containing subunit A1 p60 katanin T01G9.5 protein protein caeel Caenorhabditis elegans MNGDVQSVIRGYLERAQVAKTMSDAGRWNEAGDLLRQLMTDVKSCKISASNRDEHDARNTFLRALEANLKLVQQNVRDEDDLHEAMTRQSGSPEPPADPDVWSKPSPPLPSSSKFGATKKGVGAAGPRPREISKSTSSMSTNPADVKPANPTQGILPQNSAGDSFDASAYDAYIVQAVRGTMATNTENTMSLDDIIGMHDVKQVLHEAVTLPLLVPEFFQGLRSPWKAMVLAGPPGTGKTLIARAIASESSSTFFTVSSTDLSSKWRGDSEKIVRLLFELARFYAPSIIFIDEIDTLGGQRGNSGEHEASRRVKSEFLVQMDGSQNKFDSRRVFVLAATNIPWELDEALRRRFEKRIFIPLPDIDARKKLIEKSMEGTPKSDEINYDDLAARTEGFSGADVVSLCRTAAINVLRRYDTKSLRGGELTAAMESLKAELVRNIDFEAALQAVSPSAGPDTMLKCKEWCDSFGAM 167A3929E573CD59 egg4_caeel Inactive protein-tyrosine phosphatase egg-4 egg-4 Egg sterile protein 4 Protein-tyrosine phosphatase-like protein egg-4 T21E3.1 protein protein caeel Caenorhabditis elegans MALNSEVMFREQINAMRSQAGRKRATSLQSFCSGNTDDSSADSTDNMDMMVDYPQQKGVSCMRARFNSESTLSKSFRKKVKKLAQKDRRSKERLNGNSEEDAIEVPRGAPSTYAAPSKLRKSKALDCLVSEKPKDEGRREDSGHGADIEMAKGHFNNVRMKVFAARTAMQVEPALVMKTRKALEMKNAVLENHQSPGAFSLHAAYKIAASAESRVGSITPCNKKVTKEAMANLIRSSYDDTEITQELLFSSKFDTKWKGRYTDIYMRRDENGKKPKRPVNGQGWVMPLKSICEKFGINSTFFTNHRIDLKSARDQVLLMRLLSHDQTSTWISDIHPEAVKNETMAEYLLRELDASTMQKRVQAFKANVLADRDRVRVAGQFYNNIRIGKRMFGAARKAKYLSTIIGGMERRFEILENSVNHIPFTHSASDNNQEKCRNPRVHCKDSTRIALQFPRGQYLGDFIHANRISGKPLFNEFIMTQAPMKNTVDDFWRMVWQEEVPYIVMLTSRKEPERCEYYWPKSPSDPAVTVPGGLRIENFGVYQAPDPLFRVTHLRLIGPDREERHVEHWQGDVNNSSNMYSPLNILRLLRNASKPVVIHDHLGVSRAACLVAAEIAICSLLRGPTYKYPVQRAVQFLRQRRPFSIETPMQYIFVHRLVAFFFRDVIGSAKELDVDYERWLQERSERMFLDDLAAPIPGYRLLSPRADPDIVRMVGRPERPNYRREAPDCVGEMPNKVATVDGILSPAKSVFEF 68EAA7234FABC448 q9xtf3-2 Dual specificity tyrosine-phosphorylation-regulated kinase mbk-2 mbk-2 Dual specificity Yak1-related kinase mbk-2 Minibrain Kinase 2 F49E11.1 protein protein caeel Caenorhabditis elegans MTLFEPSTSGNRMGYRGSSNSSSGVGSGGSGSLMTQSIGGPNKHLSASHSTLNTASTHDMMHSKIPKSPSNESLSRSHTSSSGGSQGGHNSNSGSNSGFRPEDAVQTFGAKLVPFEKNEIYNYTRVFFVGSHAKKQAGVIGGANNGGYDDENGSYQLVVHDHIAYRYEVLKVIGKGSFGQVIKAFDHKYQQYVALKLVRNEKRFHRQADEEIRILDHLRRQDSDGTHNIIHMLDYFNFRNHKCITFELLSINLYELIKRNKFQGFSLMLVRKFAYSMLLCLDLLQKNRLIHCDLKPENVLLKQQGRSGIKVIDFGSSCFDDQRIYTYIQSRFYRAPEVILGTKYGMPIDMWSLGCILAELLTGYPLLPGEDENDQLALIIELLGMPPPKSLETAKRARTFITSKGYPRYCTATSMPDGSVVLAGARSKRGKMRGPPASRSWSTALKNMGDELFVDFLKRCLDWDPETRMTPAQALKHKWLRRRLPNPPRDGLESMGGLADHEVCFIIF No experimental confirmation available. 3EC0F264B291AC2C ccnb1_human G2/mitotic-specific cyclin-B1 CCNB1 CCNB protein protein human Homo sapiens Human MALRVTRNSKINAENKAKINMAGAKRVPTAPAATSKPGLRPRTALGDIGNKVSEQLQAKMPMKKEAKPSATGKVIDKKLPKPLEKVPMLVPVPVSEPVPEPEPEPEPEPVKEEKLSPEPILVDTASPSPMETSGCAPAEEDLCQAFSDVILAVNDVDAEDGADPNLCSEYVKDIYAYLRQLEEEQAVRPKYLLGREVTGNMRAILIDWLVQVQMKFRLLQETMYMTVSIIDRFMQNNCVPKKMLQLVGVTAMFIASKYEEMYPPEIGDFAFVTDNTYTKHQIRQMEMKILRALNFGLGRPLPLHFLRRASKIGEVDVEQHTLAKYLMELTMLDYDMVHFPPSQIAAGAFCLALKILDNGEWTPTLQHYLSYTEESLLPVMQHLAKNVVMVNQGLTKHMTVKNKYATSKHAKISTLPQLNSALVQDLAKAVAKV DIP protein P14635 original sequence version: 146 DIP protein P14635 original sequence version: 108 DIP protein P14635 original sequence version: 145 DIP protein P14635 original sequence version: 139 E2C4767AE8A11EC0 cdk1-mbk2-1 2 10 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 enzyme target enzyme target substrate substrate neutral component neutral component region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9XTF3-2:p.Lys196Arg Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 19:49:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys196arg' Tue Jun 30 18:20:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys196Arg' ecoli Escherichia coli (strain K12) 5 enzyme enzyme neutral component neutral component spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 3 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation hCdk-1 phosphorylates MBK-2 Fig. 2A mei1-mbk2-1 2 9 unspecified role unspecified role bait bait n-terminal 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 Q9XTF3-2:p.Ser68Ala Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:33:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser68ala' Tue Jun 30 16:11:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser68Ala' Q9XTF3-2:p.Ser68Glu mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S E Wed May 02 19:35:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser68glu' Tue Jun 30 15:44:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser68Glu' caeel Caenorhabditis elegans 7 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 n-terminal 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation S68 phosphorylation could stimulate MBK-2′s intrinsic kinase activity or MBK-2′s access to MEI-1. Fig. 3A Phosphorylation of mei-1 mei1-mbk2-2 2 9 enzyme enzyme neutral component neutral component Q9XTF3-2:p.Lys196Arg Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 16:50:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys196arg' Tue Jun 30 17:15:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys196Arg' n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 7 enzyme target enzyme target substrate substrate neutral component neutral component n-terminal 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. 3B egg3-mbk2-1 1 9 unspecified role unspecified role bait bait n-terminal 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 ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association To test whether MBK-2 also binds to EGG-4 and EGG-5, we expressed each separately as GST and FLAG fusion proteins in E. coli. GST:MBK-2-coupled beads could pull down FLAG:EGG-4 and FLAG:EGG-5 (and FLAG:EGG-3) from E. coli extracts (Figure 5A). Fig. 5A egg4-mbk2-1 1 9 unspecified role unspecified role bait bait n-terminal 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 Q9XTF3-2:p.Lys196Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 16:41:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys196arg' Tue Jun 30 17:02:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys196Arg' Q9XTF3-2:p.[Tyr325Phe;Tyr327Phe] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 15:05:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr325phe,tyr327phe' Tue Jun 30 17:30:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr325Phe;Tyr327Phe]' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association To test whether MBK-2 also binds to EGG-4 and EGG-5, we expressed each separately as GST and FLAG fusion proteins in E. coli. GST:MBK-2-coupled beads could pull down FLAG:EGG-4 and FLAG:EGG-5 (and FLAG:EGG-3) from E. coli extracts (Figure 5A). Fig. 5A egg5-mbk2-1 1 9 unspecified role unspecified role bait bait n-terminal 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 Q9XTF3-2:p.Lys196Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 19:24:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys196arg' Tue Jun 30 15:36:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys196Arg' Q9XTF3-2:p.[Tyr325Phe;Tyr327Phe] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F certain certain sequence position certain certain certain sequence position certain Y F Fri Jul 22 10:53:59 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr327phe,tyr325phe' Wed May 02 18:41:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr325phe,tyr327phe' Tue Jun 30 19:50:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr325Phe;Tyr327Phe]' ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association To test whether MBK-2 also binds to EGG-4 and EGG-5, we expressed each separately as GST and FLAG fusion proteins in E. coli. GST:MBK-2-coupled beads could pull down FLAG:EGG-4 and FLAG:EGG-5 (and FLAG:EGG-3) from E. coli extracts (Figure 5A). Fig. 5A