#ID(s) interactor A	ID(s) interactor B	Alt. ID(s) interactor A	Alt. ID(s) interactor B	Alias(es) interactor A	Alias(es) interactor B	Interaction detection method(s)	Publication 1st author(s)	Publication Identifier(s)	Taxid interactor A	Taxid interactor B	Interaction type(s)	Source database(s)	Interaction identifier(s)	Confidence value(s)	Expansion method(s)	Biological role(s) interactor A	Biological role(s) interactor B	Experimental role(s) interactor A	Experimental role(s) interactor B	Type(s) interactor A	Type(s) interactor B	Xref(s) interactor A	Xref(s) interactor B	Interaction Xref(s)	Annotation(s) interactor A	Annotation(s) interactor B	Interaction annotation(s)	Host organism(s)	Interaction parameter(s)	Creation date	Update date	Checksum(s) interactor A	Checksum(s) interactor B	Interaction Checksum(s)	Negative	Feature(s) interactor A	Feature(s) interactor B	Stoichiometry(s) interactor A	Stoichiometry(s) interactor B	Identification method participant A	Identification method participant B
uniprotkb:P49814	uniprotkb:P39126	intact:EBI-7827708|ensemblbacteria:CAB14872|intact:MINT-8365164	intact:EBI-7829570|ensemblbacteria:CAB14873|intact:MINT-1711252	psi-mi:mdh_bacsu(display_long)|uniprotkb:mdh(gene name)|psi-mi:mdh(display_short)|uniprotkb:Vegetative protein 69(gene name synonym)|uniprotkb:citH(gene name synonym)|uniprotkb:BSU29120(locus name)	psi-mi:idh_bacsu(display_long)|uniprotkb:icd(gene name)|psi-mi:icd(display_short)|uniprotkb:citC(gene name synonym)|uniprotkb:BSU29130(locus name)|uniprotkb:Oxalosuccinate decarboxylase(gene name synonym)|uniprotkb:"NADP(+)-specific ICDH"(gene name synonym)|uniprotkb:IDP(gene name synonym)	psi-mi:"MI:0107"(surface plasmon resonance)	Bartholomae et al. (2013)	imex:IM-22019|pubmed:24325460	taxid:224308(bacsu)|taxid:224308(Bacillus subtilis)	taxid:224308(bacsu)|taxid:224308(Bacillus subtilis)	psi-mi:"MI:0407"(direct interaction)	psi-mi:"MI:0471"(MINT)	intact:EBI-9028685|imex:IM-22019-3	-	-	psi-mi:"MI:0499"(unspecified role)	psi-mi:"MI:0499"(unspecified role)	psi-mi:"MI:0496"(bait)	psi-mi:"MI:0498"(prey)	psi-mi:"MI:0326"(protein)	psi-mi:"MI:0326"(protein)	ensemblbacteria:BSU_29120(gene)|ensemblbacteria:CAB14872(transcript)|go:"GO:0005737"(cytoplasm)|go:"GO:0005975"(carbohydrate metabolic process)|go:"GO:0006099"(tricarboxylic acid cycle)|go:"GO:0019752"(carboxylic acid metabolic process)|go:"GO:0030060"(L-malate dehydrogenase activity)|interpro:IPR015955(Lactate dehydrogenase/glycoside hydrolase, family 4, C-terminal)|interpro:IPR022383|interpro:IPR011275(Malate dehydrogenase, NAD-dependent)|refseq:NP_390790.1|interpro:IPR001557(L-lactate/malate dehydrogenase)|interpro:IPR001236(Lactate/malate dehydrogenase)|refseq:WP_003229437.1|interpro:IPR036291	interpro:IPR004439(Isocitrate dehydrogenase NADP-dependent, prokaryotic)|ensemblbacteria:BSU_29130(gene)|ensemblbacteria:CAB14873(transcript)|go:"GO:0000287"(magnesium ion binding)|go:"GO:0004450"("isocitrate dehydrogenase (NADP+) activity")|go:"GO:0006097"(glyoxylate cycle)|go:"GO:0006099"(tricarboxylic acid cycle)|go:"GO:0051287"(NAD binding)|refseq:WP_003229433.1|interpro:IPR019818|rcsb pdb:1HQS|interpro:IPR024084|refseq:NP_390791.1	-	comment:mint	comment:mint	figure legend:f4|comment:"the interaction of  Mdh and Icd was characterized in the presence of their substrates and products as well.  Therefore, isocitrate, α-ketoglutarate, malate, and oxaloacetate, as well as the cofactors Mg2+,  NADP+, and NADPH for Icd, and NAD+ and NADH for Mdh were added alone, in double  and triple combinations. The concentrations of substrates and cofactors were adjusted to the  respective intracellular concentrations which were determined for B. subtilis and E. coli in the  exponential growth phase and at high availability of carbon and inorganic nitrogen sources,  respectively [20-23]. The sensorgrams of these SPR analyses are shown in Figure 2. The  single substances and their double combinations or the triple combinations of Mg2+, α-  ketoglutarate and NADPH, of Mg2+, malate and NAD+ and of Mg2+, oxaloacetate and NADH  did not show significant changes of the sensorgram for 20 μM Icd. Only the combination of  substrate and cofactors of Icd, isocitrate, NADP+ and Mg2+, stimulated the interaction of Mdh  and Icd to a high level (Figure 2). Thus, the stimulatory effect of the triple combination of the  substrates and cofactors of Icd is highly specific."|comment:"The interaction of IcdS104P with Mdh was analysed by SPR in the presence or the absence of  substrates, cofactors and products, and their double or triple combinations as described before.  Again only the triple combination of 1 mM isocitrate, 0.5 mM NADP+ and 5 mM MgCl2  stimulated binding of the IcdS104P mutant to Mdh and, therefore, this mutant was a  promising candidate for a quantitative analysis of the interaction. Interestingly, the  sensorgrams of the analysis of IcdS104P with Mdh without substrates showed a stronger  interaction than wildtype Icd (Figure 4). The kinetic analysis revealed an association rate  constant (ka) of 2.0±0.2•102 M-1 s-1, a dissociation rate constant (kd) of 1.0±0.1•10-3 s-1 and an  equilibrium constant (KD) of 5.0±0.1 μM (χ²=1.8-4.8). For the kinetic analysis of the  IcdS104P-Mdh interaction in the presence of substrates and cofactors, 0.5 to 20 μM IcdS104P  were injected with 1 mM isocitrate, 0.5 mM NADP+ and 5 mM MgCl2 on a Mdh-coated CM5  chip. The kinetic analysis revealed an association rate constant (ka) of 1.7±0.7•103 M-1 s-1, a  dissociation rate constant (kd) of 2.6±0.6•10-4 s-1 and an equilibrium constant (KD) of  150±1 nM (χ²=2.3-3.0). The comparison of the quantitative data shows a 33-fold stimulation  of the interaction of IcdS104P, and the KD values are in a reasonable range for protein-protein  interactions."|full coverage:Only protein-protein interactions|curation depth:imex curation	taxid:-1(in vitro)|taxid:-1(In vitro)	kd:150.0x10^-9(molar)	2013/12/04	2014/10/16	rogid:eNUWZhidR10KBy7Yuou7/Lf0jE0224308	rogid:pK5DkW3/QjuGic0SQ2zMNsQgyWQ224308	intact-crc:ACA8378E3E623AB5|rigid:loKqZS/BY7HXOJ6n7v1aIvY+4l0	false	his tag:?-?	mutation increasing interaction:104-104|his tag:?-?	-	-	psi-mi:"MI:0396"(predetermined participant)	psi-mi:"MI:0396"(predetermined participant)
uniprotkb:P49814	uniprotkb:P39126	intact:EBI-7827708|ensemblbacteria:CAB14872|intact:MINT-8365164	intact:EBI-7829570|ensemblbacteria:CAB14873|intact:MINT-1711252	psi-mi:mdh_bacsu(display_long)|uniprotkb:mdh(gene name)|psi-mi:mdh(display_short)|uniprotkb:Vegetative protein 69(gene name synonym)|uniprotkb:citH(gene name synonym)|uniprotkb:BSU29120(locus name)	psi-mi:idh_bacsu(display_long)|uniprotkb:icd(gene name)|psi-mi:icd(display_short)|uniprotkb:citC(gene name synonym)|uniprotkb:BSU29130(locus name)|uniprotkb:Oxalosuccinate decarboxylase(gene name synonym)|uniprotkb:"NADP(+)-specific ICDH"(gene name synonym)|uniprotkb:IDP(gene name synonym)	psi-mi:"MI:0107"(surface plasmon resonance)	Bartholomae et al. (2013)	imex:IM-22019|pubmed:24325460	taxid:224308(bacsu)|taxid:224308(Bacillus subtilis)	taxid:224308(bacsu)|taxid:224308(Bacillus subtilis)	psi-mi:"MI:0407"(direct interaction)	psi-mi:"MI:0471"(MINT)	intact:EBI-9028643|imex:IM-22019-1	-	-	psi-mi:"MI:0499"(unspecified role)	psi-mi:"MI:0499"(unspecified role)	psi-mi:"MI:0496"(bait)	psi-mi:"MI:0498"(prey)	psi-mi:"MI:0326"(protein)	psi-mi:"MI:0326"(protein)	ensemblbacteria:BSU_29120(gene)|ensemblbacteria:CAB14872(transcript)|go:"GO:0005737"(cytoplasm)|go:"GO:0005975"(carbohydrate metabolic process)|go:"GO:0006099"(tricarboxylic acid cycle)|go:"GO:0019752"(carboxylic acid metabolic process)|go:"GO:0030060"(L-malate dehydrogenase activity)|interpro:IPR015955(Lactate dehydrogenase/glycoside hydrolase, family 4, C-terminal)|interpro:IPR022383|interpro:IPR011275(Malate dehydrogenase, NAD-dependent)|refseq:NP_390790.1|interpro:IPR001557(L-lactate/malate dehydrogenase)|interpro:IPR001236(Lactate/malate dehydrogenase)|refseq:WP_003229437.1|interpro:IPR036291	interpro:IPR004439(Isocitrate dehydrogenase NADP-dependent, prokaryotic)|ensemblbacteria:BSU_29130(gene)|ensemblbacteria:CAB14873(transcript)|go:"GO:0000287"(magnesium ion binding)|go:"GO:0004450"("isocitrate dehydrogenase (NADP+) activity")|go:"GO:0006097"(glyoxylate cycle)|go:"GO:0006099"(tricarboxylic acid cycle)|go:"GO:0051287"(NAD binding)|refseq:WP_003229433.1|interpro:IPR019818|rcsb pdb:1HQS|interpro:IPR024084|refseq:NP_390791.1	-	comment:mint	comment:mint	figure legend:f1 f2|comment:"the interaction of  Mdh and Icd was characterized in the presence of their substrates and products as well.  Therefore, isocitrate, α-ketoglutarate, malate, and oxaloacetate, as well as the cofactors Mg2+,  NADP+, and NADPH for Icd, and NAD+ and NADH for Mdh were added alone, in double  and triple combinations. The concentrations of substrates and cofactors were adjusted to the  respective intracellular concentrations which were determined for B. subtilis and E. coli in the  exponential growth phase and at high availability of carbon and inorganic nitrogen sources,  respectively [20-23]. The sensorgrams of these SPR analyses are shown in Figure 2. The  single substances and their double combinations or the triple combinations of Mg2+, α-  ketoglutarate and NADPH, of Mg2+, malate and NAD+ and of Mg2+, oxaloacetate and NADH  did not show significant changes of the sensorgram for 20 μM Icd. Only the combination of  substrate and cofactors of Icd, isocitrate, NADP+ and Mg2+, stimulated the interaction of Mdh  and Icd to a high level (Figure 2). Thus, the stimulatory effect of the triple combination of the  substrates and cofactors of Icd is highly specific."|full coverage:Only protein-protein interactions|curation depth:imex curation	taxid:-1(in vitro)|taxid:-1(In vitro)	-	2013/12/04	2014/10/16	rogid:eNUWZhidR10KBy7Yuou7/Lf0jE0224308	rogid:pK5DkW3/QjuGic0SQ2zMNsQgyWQ224308	intact-crc:DDA849615932EBF0|rigid:loKqZS/BY7HXOJ6n7v1aIvY+4l0	false	his tag:?-?	his tag:?-?	-	-	psi-mi:"MI:0396"(predetermined participant)	psi-mi:"MI:0396"(predetermined participant)
uniprotkb:P49814	uniprotkb:P39126	intact:EBI-7827708|ensemblbacteria:CAB14872|intact:MINT-8365164	intact:EBI-7829570|ensemblbacteria:CAB14873|intact:MINT-1711252	psi-mi:mdh_bacsu(display_long)|uniprotkb:mdh(gene name)|psi-mi:mdh(display_short)|uniprotkb:Vegetative protein 69(gene name synonym)|uniprotkb:citH(gene name synonym)|uniprotkb:BSU29120(locus name)	psi-mi:idh_bacsu(display_long)|uniprotkb:icd(gene name)|psi-mi:icd(display_short)|uniprotkb:citC(gene name synonym)|uniprotkb:BSU29130(locus name)|uniprotkb:Oxalosuccinate decarboxylase(gene name synonym)|uniprotkb:"NADP(+)-specific ICDH"(gene name synonym)|uniprotkb:IDP(gene name synonym)	psi-mi:"MI:0030"(cross-linking study)	Bartholomae et al. (2013)	imex:IM-22019|pubmed:24325460	taxid:224308(bacsu)|taxid:224308(Bacillus subtilis)	taxid:224308(bacsu)|taxid:224308(Bacillus subtilis)	psi-mi:"MI:0915"(physical association)	psi-mi:"MI:0471"(MINT)	intact:EBI-9028674|imex:IM-22019-2	-	-	psi-mi:"MI:0499"(unspecified role)	psi-mi:"MI:0499"(unspecified role)	psi-mi:"MI:0498"(prey)	psi-mi:"MI:0496"(bait)	psi-mi:"MI:0326"(protein)	psi-mi:"MI:0326"(protein)	ensemblbacteria:BSU_29120(gene)|ensemblbacteria:CAB14872(transcript)|go:"GO:0005737"(cytoplasm)|go:"GO:0005975"(carbohydrate metabolic process)|go:"GO:0006099"(tricarboxylic acid cycle)|go:"GO:0019752"(carboxylic acid metabolic process)|go:"GO:0030060"(L-malate dehydrogenase activity)|interpro:IPR015955(Lactate dehydrogenase/glycoside hydrolase, family 4, C-terminal)|interpro:IPR022383|interpro:IPR011275(Malate dehydrogenase, NAD-dependent)|refseq:NP_390790.1|interpro:IPR001557(L-lactate/malate dehydrogenase)|interpro:IPR001236(Lactate/malate dehydrogenase)|refseq:WP_003229437.1|interpro:IPR036291	interpro:IPR004439(Isocitrate dehydrogenase NADP-dependent, prokaryotic)|ensemblbacteria:BSU_29130(gene)|ensemblbacteria:CAB14873(transcript)|go:"GO:0000287"(magnesium ion binding)|go:"GO:0004450"("isocitrate dehydrogenase (NADP+) activity")|go:"GO:0006097"(glyoxylate cycle)|go:"GO:0006099"(tricarboxylic acid cycle)|go:"GO:0051287"(NAD binding)|refseq:WP_003229433.1|interpro:IPR019818|rcsb pdb:1HQS|interpro:IPR024084|refseq:NP_390791.1	-	comment:mint	comment:mint	figure legend:f3|comment:As shown in Figure 3, FLAG-Mdh stability was not affected by  formaldehyde treatment because equal amounts of Mdh were detected in the crude extracts  before purification of Strep-Icd or Strep-IcdS104P. Small amounts of the FLAG-tagged Mdh  were detected in the fractions from non-cross-linked cultures when the Strep-tagged wildtype  and the Icd mutant variant served as the bait proteins. In contrast, the amount of co-purified  FLAG-Mdh was increased when both wildtype Strep-Icd and Strep-IcdS104P were purified  from the cross-linked cells. Thus, mutant and wildtype Icd show the same interaction pattern  with Mdh in vivo and we concluded that the mutation of serine 104 to proline in Icd did not  affect its interaction with Mdh.|full coverage:Only protein-protein interactions|curation depth:imex curation	taxid:-1(in vitro)|taxid:-1(In vitro)	-	2013/12/04	2014/10/16	rogid:eNUWZhidR10KBy7Yuou7/Lf0jE0224308	rogid:pK5DkW3/QjuGic0SQ2zMNsQgyWQ224308	intact-crc:19782333A0B1CC49|rigid:loKqZS/BY7HXOJ6n7v1aIvY+4l0	false	flag tag:?-?	strep tag:?-?	-	-	psi-mi:"MI:0705"(anti tag western blot)	psi-mi:"MI:0396"(predetermined participant)