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/ Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. J. Mol. Biol. (0022-2836) 2003 imex curation Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro aut-page aut-page weight by staining molecular weight estimation by staining weight by staining Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr Accepted 2013-AUG-08 by PPORRAS 09-Sep-2013: Contacted by IntAct-Help. J. Mol. Biol. (0022-2836) 2003 Only protein-protein interactions imex curation Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. J. Mol. Biol. (0022-2836) 2003 imex curation Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro dsc differential scanning calorimetry dsc predetermined predetermined participant predetermined Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr Accepted 2013-AUG-08 by PPORRAS 09-Sep-2013: Contacted by IntAct-Help. J. Mol. Biol. (0022-2836) 2003 Only protein-protein interactions imex curation Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. J. Mol. Biol. (0022-2836) 2003 imex curation Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa predetermined predetermined participant predetermined Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr Accepted 2013-AUG-08 by PPORRAS 09-Sep-2013: Contacted by IntAct-Help. J. Mol. Biol. (0022-2836) 2003 Only protein-protein interactions imex curation Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. J. Mol. Biol. (0022-2836) 2003 imex curation Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr Accepted 2013-AUG-08 by PPORRAS 09-Sep-2013: Contacted by IntAct-Help. J. Mol. Biol. (0022-2836) 2003 Only protein-protein interactions imex curation Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. Evidence of a thermal unfolding dimeric intermediate for the Escherichia coli histone-like HU proteins: thermodynamics and structure. J. Mol. Biol. (0022-2836) 2003 imex curation Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro cd circular dichroism CD CD cd predetermined predetermined participant predetermined Ramstein J., Hervouet N., Coste F., Zelwer C., Oberto J., Castaing B. castaing@cnrs-orleans.fr Accepted 2013-AUG-08 by PPORRAS 09-Sep-2013: Contacted by IntAct-Help. J. Mol. Biol. (0022-2836) 2003 Only protein-protein interactions imex curation dbhb_ecoli DNA-binding protein HU-beta hupB hopD b0440 NS1 HU-1 JW0430 protein protein ecoli Escherichia coli (strain K12) MNKSQLIDKIAAGADISKAAAGRALDAIIASVTESLKEGDDVALVGFGTFAVKERAARTGRNPQTGKEITIAAAKVPSFRAGKALKDAVN DIP protein P0ACF4 original sequence version: 74 DIP protein P0ACF4 original sequence version: 43 C56A3C766D53BB96 dbha_ecoli DNA-binding protein HU-alpha hupA b4000 NS2 HU-2 JW3964 protein protein ecoli Escherichia coli (strain K12) MNKTQLIDVIAEKAELSKTQAKAALESTLAAITESLKEGDAVQLVGFGTFKVNHRAERTGRNPQTGKEIKIAAANVPAFVSGKALKDAVK 6B5B8536FDBD13DC nicked-dna-12875839-forw nicked DNA from pmid:12875839 forward ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) TACGGATCGCAGCGGTTAGGGAAGTTGG nicked-dna-12875839-rev nicked DNA from pmid:12875839 reverse ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) ATGCCTAGCGTCGACCCAATCCCTTCAACC hupa-hupb-1 1 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1c hupa-hupb-2 4 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 1a, 2 hupa-hupb-3 2 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3, 5, 6, Table 1 Higher protein concentration shifted the thermodynamic profile slightly to the right, higher ionic strength caused a large shift to the right. hupa-1 2 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3, 4, 5, 6, Table 1 Higher protein concentration shifted the thermodynamic profile slightly to the right, higher ionic strength caused a large shift to the right. hupb-1 2 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3, 5, 6, Table 1 Higher protein concentration shifted the thermodynamic profile slightly to the right, higher ionic strength caused a large shift to the right. However, the shift for peak 2 is smaller than that observed for the dbha/b heterodimer (EBI-6678417) or dbha homodimer (EBI-6678436). hupa-hupb-4 5 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Table 2 hupb-2 5 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Table 2 hupa-2 5 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 7, Table 2 hupa-hupb-5 3 8 unspecified role unspecified role bait bait dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain g14-phospho phosphorus containing modified residue phosphorus containing modified residue certain certain sequence position certain certain certain sequence position certain 2 bases (TG) were removed between C13 and G14 compared to the reverse strand. chemical synthesis Chemical synthesis (Chemical synthesis) 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain c30-radiophosph 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 41 36 physical association physical association Table 3 dK values given for interaction at 20C, other values are: 15 nM at 5C and 15C. hupa-3 3 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait g14-phospho phosphorus containing modified residue phosphorus containing modified residue certain certain sequence position certain certain certain sequence position certain 2 bases (TG) were removed between C13 and G14 compared to the reverse strand. dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 9 unspecified role unspecified role bait bait dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain c30-radiophosph 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 50 48 physical association physical association Table 3 dK values given for interaction at 20C, other values are: 10 nM at 5C and 15 nM 15C. hupb-3 3 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait c30-radiophosph 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 8 unspecified role unspecified role bait bait dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain g14-phospho phosphorus containing modified residue phosphorus containing modified residue certain certain sequence position certain certain certain sequence position certain 2 bases (TG) were removed between C13 and G14 compared to the reverse strand. chemical synthesis Chemical synthesis (Chemical synthesis) 57 59 physical association physical association Table 3 dK values given for interaction at 20C, other values are: 40 nM at 5C and 70 nM 15C.