MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. FEBS J. (1742-464X) 2011 imex curation Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. Only protein-protein interactions in vitro In vitro crosslink cross-linking study crosslink predetermined predetermined participant predetermined Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. FEBS J. (1742-464X) 2011 Only protein-protein interactions imex curation Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. FEBS J. (1742-464X) 2011 imex curation Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. FEBS J. (1742-464X) 2011 Only protein-protein interactions imex curation Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. FEBS J. (1742-464X) 2011 imex curation Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. FEBS J. (1742-464X) 2011 Only protein-protein interactions imex curation Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis. FEBS J. (1742-464X) 2011 imex curation Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. Only protein-protein interactions bacsu Bacillus subtilis anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Kayumov A., Heinrich A., Fedorova K., Ilinskaya O., Forchhammer K. FEBS J. (1742-464X) 2011 Only protein-protein interactions imex curation gln1a_bacsu Glutamine synthetase Glutamine synthetase I alpha BSU17460 Glutamate--ammonia ligase glnA protein protein bacsu Bacillus subtilis MAKYTREDIEKLVKEENVKYIRLQFTDILGTIKNVEIPVSQLGKALDNKVMFDGSSIEGFVRIEESDMYLYPDLNTFVIFPWTAEKGKVARFICDIYNPDGTPFEGDPRNNLKRILKEMEDLGFSDFNLGPEPEFFLFKLDEKGEPTLELNDKGGYFDLAPTDLGENCRRDIVLELEEMGFEIEASHHEVAPGQHEIDFKYAGAVRSCDDIQTFKLVVKTIARKHGLHATFMPKPLFGVNGSGMHCNLSLFKNGVNAFFDENADLQLSETAKHFIAGIVKHATSFTAVTNPTVNSYKRLVPGYEAPCYVAWSAQNRSPLIRIPASRGISTRVEVRSVDPAANPYLALSVLLAAGLDGIKNKLEAPAPIDRNIYVMSKEERMENGIVDLPATLAEALEEFKSNEVMVKALGEHLFEHFIEAKEIEWDMFRTQVHPWEREQYMSQY 83A5657CE1388AB0 83A5657CE1388AB0 glnk_bacsu Sensor histidine kinase GlnK glnK ycbA yzgA BSU02440 nrgB protein protein bacsu Bacillus subtilis MLITVPLAGELKFYPLNEEFRVSFGAPVFFFFLSLLRHVPAVLPGFLTGAAVFIFRVFLELWGGGHNGLTPILYDQASGFFFYMTYACLFSILKANRFRERPIMLGFIGFMIEVVSDCVELTVQFLIFHTVVTPEKITDIAVIAISHTFIVMSFYSVLKLYETQSREKQTRQQHEHMLMIVSNLYEETVHLKKTLKTTEKVTNDSYQLYREMKGKDVQLSGRILRLAGEIHEVKKDNQRIFAGLSKLISNESLRDYMRASDLLQLVIRMNEKYAEALGKQIDFYCSIEGEHDEYHVFIVLSIINNLTANAVEAMDEEGMVSLRLRKPNESMVEFQVEDNGPGISEKIGDIVFDPGFTSKYDEFGTPSTGIGLSYVKEIVTELEGDITFDNQQRGVVFAIRLPVRHLIQKG mint 88D0E033366113E2 tnra_bacsu HTH-type transcriptional regulator TnrA tnrA scgR BSU13310 Transcriptional nitrogen regulatory protein A protein protein bacsu Bacillus subtilis MTTEDHSYKDKKVISIGIVSELTGLSVRQIRYYEERKLIYPQRSSRGTRKYSFADVERLMDIANKREDGVQTAEILKDMRKKEQMLKNDPQVRKKMLEGQLNAHFRYKNR mint Regulator of transcription that activates or represses many genes during nitrogen-limited growth; among others, it activates nrgAB, nasABCDE, gabP, and represses glnRA 201AA41E76E0E631 tnra-glnk 4 tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level glnK 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1a mint When the cells were shifted to nitrate-deprived medium prior to 25 extraction of the proteins, much less TnrA was immunoprecipitated, and in consequence, also less 26 GlnK was detected, since TnrA is degraded by proteolysis following the shift to nitrate-deprived 27 medium [15]. By contrast, in the AmtB-deficient mutant strain, the same amount of TnrA was 28 immunoprecipitated and the same amount of GlnK was co-purified with TnrA in both nitrogen 29 regimes (Fig. 1A, lanes I and II). tnra-glna-2 4 tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level GS 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1a mint In the TnrA-immunoprecipitate of GlnK-deficient cells,GS was co-purified with TnrA and the recovery of GS was independent on whether the cells were nitrate-grown or shifted to nitrate-deprived medium. glnk-tnra-1 4 glnK 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1b mint TnrA was co-purified with immunoprecipitated GlnK from extracts of both wild type and AmtB deficient cells (Fig. 1B) glna-tnra-1 4 tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level GS 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1c mint TnrA was co-purified with immunoprecipitated GlnK was recovered by GS immunoprecipitation only in the GlnK-deficient mutant (Fig. 1C). tnra-glnk-1 3 tnrA 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain his tag 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 glnK 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction mint Fig. 2A shows a response-difference sensorgram (FC2-FC1) of interactions of 18 GlnK with 19 immobilized TnrA. For this analysis, an analyte concentration of 40 nM GlnK (trimer) was used. A, ATP effect on dissociation of the TnrA-GlnK complex. First, GlnK (40 nM trimers) was injected to the His6-TnrA surface. After 50 s of washing with HBS 16 buffer, 25 μl of 2 mM ATP was injected (indicated by the arrow), which removed the GlnK bound to the His6-TnrA surface within a few seconds. B, Binding of GlnK to TnrA in the presence of different Mg2+ or Mn2+ concentrations with or without 2 mM ATP and 1 mM 2-OG present, as indicated. Fig. 3. Influence of various effector molecules on the interaction of GlnK with the His6- TnrA surface f2 f3 f6a tnra-glna-3 3 tnrA 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified his tag 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain GS 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction mint The GS-TnrA complex was quite stable under the conditions used: almost no complex dissociation appeared after the injection phase. In contrast to GlnK, no efficient effector molecule was found to remove GS from the His6-TnrA surface. 10 mM ATP caused only partial release of GS from TnrA (Fig 5A). the C-terminus of TnrA is absolutely required for GS binding: deletion of already 6 amino acids drastically abolished this interaction (Fig. 6 A, B). f5 f6a tnra-glna-4 2 tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain his tag 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 GS 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f6b mint glna-tnra-2 2 tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain GS 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f6b mint tnra-glnk-2 2 glnK 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain his tag 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 direct interaction direct interaction f6c mint glnk-tnra-2 2 glnK 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain his tag 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 direct interaction direct interaction f6c mint tnra-3 1 tnrA 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction sf3 mint