mpidb The Microbial Protein Interaction Database cunningham-2009-1 The histidine kinase inhibitor Sda binds near the site of autophosphorylation and may sterically hinder autophosphorylation and phosphotransfer to Spo0F. in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Cunningham K.A., Burkholder W.F. Mol. Microbiol. (0950-382X) 2009 Only protein-protein interactions imex curation cunningham-2009-5 The histidine kinase inhibitor Sda binds near the site of autophosphorylation and may sterically hinder autophosphorylation and phosphotransfer to Spo0F. in vitro In vitro protein kinase assay protein kinase assay tag visualisation tag visualisation Cunningham K.A., Burkholder W.F. C-terminal tag is radio-labelled via a PKA-dependent phosphorylation assay. Mol. Microbiol. (0950-382X) 2009 Only protein-protein interactions imex curation cunningham-2009-6 The histidine kinase inhibitor Sda binds near the site of autophosphorylation and may sterically hinder autophosphorylation and phosphotransfer to Spo0F. in vitro In vitro phosphotransferase phosphotransferase assay phosphotransferase weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Cunningham K.A., Burkholder W.F. C-terminal tag is radio-labelled via a PKA-dependent phosphorylation assay. Mol. Microbiol. (0950-382X) 2009 Only protein-protein interactions imex curation cunningham-2009-2 The histidine kinase inhibitor Sda binds near the site of autophosphorylation and may sterically hinder autophosphorylation and phosphotransfer to Spo0F. in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Cunningham K.A., Burkholder W.F. Mol. Microbiol. (0950-382X) 2009 Only protein-protein interactions imex curation cunningham-2009-3 The histidine kinase inhibitor Sda binds near the site of autophosphorylation and may sterically hinder autophosphorylation and phosphotransfer to Spo0F. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Cunningham K.A., Burkholder W.F. C-terminal tag is radio-labelled via a PKA-dependent phosphorylation assay. Mol. Microbiol. (0950-382X) 2009 Only protein-protein interactions imex curation cunningham-2009-4 The histidine kinase inhibitor Sda binds near the site of autophosphorylation and may sterically hinder autophosphorylation and phosphotransfer to Spo0F. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Cunningham K.A., Burkholder W.F. Mol. Microbiol. (0950-382X) 2009 Only protein-protein interactions imex curation kina_bacsu Sporulation kinase A kinA Stage II sporulation protein F Stage II sporulation protein J gsiC scoD spoIIF spoIIJ BSU13990 protein protein bacsu Bacillus subtilis MEQDTQHVKPLQTKTDIHAVLASNGRIIYISANSKLHLGYLQGEMIGSFLKTFLHEEDQFLVESYFYNEHHLMPCTFRFIKKDHTIVWVEAAVEIVTTRAERTEREIILKMKVLEEETGHQSLNCEKHEIEPASPESTTYITDDYERLVENLPSPLCISVKGKIVYVNSAMLSMLGAKSKDAIIGKSSYEFIEEEYHDIVKNRIIRMQKGMEVGMIEQTWKRLDGTPVHLEVKASPTVYKNQQAELLLLIDISSRKKFQTILQKSRERYQLLIQNSIDTIAVIHNGKWVFMNESGISLFEAATYEDLIGKNIYDQLHPCDHEDVKERIQNIAEQKTESEIVKQSWFTFQNRVIYTEMVCIPTTFFGEAAVQVILRDISERKQTEELMLKSEKLSIAGQLAAGIAHEIRNPLTAIKGFLQLMKPTMEGNEHYFDIVFSELSRIELILSELLMLAKPQQNAVKEYLNLKKLIGEVSALLETQANLNGIFIRTSYEKDSIYINGDQNQLKQVFINLIKNAVESMPDGGTVDIIITEDEHSVHVTVKDEGEGIPEKVLNRIGEPFLTTKEKGTGLGLMVTFNIIENHQGVIHVDSHPEKGTAFKISFPKK A8BB97FE844B32EE sda_bacsu Sporulation inhibitor sda sda Histidine kinase KinA inhibitor BSU25690 protein protein bacsu Bacillus subtilis MNWVPSMRKLSDELLIESYFKATEMNLNRDFIELIENEIKRRSLGHIISVSS sp0f_bacsu Sporulation initiation phosphotransferase F spo0F Stage 0 sporulation protein F BSU37130 protein protein bacsu Bacillus subtilis MMNEKILIVDDQYGIRILLNEVFNKEGYQTFQAANGLQALDIVTKERPDLVLLDMKIPGMDGIEILKRMKVIDENIRVIIMTAYGELDMIQESKELGALTHFAKPFDIDEIRDAVKKYLPLKSN 4130377E3558D698 kina-kina-3 1334784 n/a 1334790 enzyme enzyme unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli not-defined-1047097060 1334790 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli not-defined-2003158388 1334791 inhibitor inhibitor unspecified role unspecified role physical association physical association false false false Fig. 1, 2B Higher concentrations of Sda are required to inhibit the autophosphorylation activities of KinA(P410L) and KinA(P410L/F436S) than to inhibit wild-type KinA kina-kina-4 1334784 not-defined-1047097060 1334790 enzyme target enzyme target substrate substrate unspecified role unspecified role region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli n/a 1334790 enzyme enzyme unspecified role unspecified role region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli phosphorylation phosphorylation reaction phosphorylation false false false Fig. 1 kina 1334785 n/a 1334790 putative self putative self putative self putative self n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal rrasv tag tag tag insertion analysis insertion analysis certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli phosphorylation phosphorylation reaction phosphorylation false true false Fig. 8 kina-sda-4 1334785 n/a 1334790 putative self putative self putative self putative self n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal rrasv tag tag tag insertion analysis insertion analysis certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli n/a 1334791 inhibitor inhibitor unspecified role unspecified role direct interaction direct interaction false false false Fig. 8 kina-spo0f 1334786 n/a 1334792 phosphate acceptor phosphate acceptor unspecified role unspecified role c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecolx Escherichia coli n/a 1334790 photon donor photon donor photon donor unspecified role unspecified role region 32p radiolabel 32p radiolabel P32 32P 32P P32 insertion analysis insertion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecolx Escherichia coli phosphotransfer phosphotransfer reaction phosphotransfer false false false Fig. 10 kina-sda-5 1334786 n/a 1334791 inhibitor inhibitor unspecified role unspecified role ecolx Escherichia coli n/a 1334792 phosphate acceptor phosphate acceptor unspecified role unspecified role c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecolx Escherichia coli n/a 1334790 photon donor photon donor photon donor unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 32p radiolabel 32p radiolabel P32 32P 32P P32 insertion analysis insertion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli physical association physical association false false false Fig. 10 kina-sda 1334787 not-defined-2003158388 1334791 unspecified role unspecified role prey prey n/a 1334790 unspecified role unspecified role bait bait P16497:p.[Pro410Leu;Phe436Ser] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P16497:p.Pro410Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false Fig. 2A kina-sda-1 1334788 n/a 1334790 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal rrasv tag tag tag insertion analysis insertion analysis certain certain sequence position certain certain certain sequence position certain false P16497:p.Tyr431Cys mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli not-defined-2003158388 1334791 unspecified role unspecified role prey prey direct interaction direct interaction false false false Fig. 4, 9 Sda Ser12Cys substitution restored the interaction of Sda with KinA(Tyr431Cys) kina-sda-3 1334789 not-defined-2003158388 1334791 unspecified role unspecified role neutral component neutral component n/a 1334790 unspecified role unspecified role neutral component neutral component P16497:p.Tyr431Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P16497:p.Ala413Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false Fig. 4