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/ Hydrolysis of the 5'-p-nitrophenyl ester of TMP by the proofreading exonuclease (epsilon) subunit of Escherichia coli DNA polymerase III. Hydrolysis of the 5'-p-nitrophenyl ester of TMP by the proofreading exonuclease (epsilon) subunit of Escherichia coli DNA polymerase III. Biochemistry (0006-2960) 2002 imex curation Hamdan S., Bulloch E.M., Thompson P.R., Beck J.L., Yang J.Y., Crowther J.A., Lilley P.E., Carr P.D., Ollis D.L., Brown S.E., Dixon N.E. dixon@rsc.anu.edu.au 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Hamdan S., Bulloch E.M., Thompson P.R., Beck J.L., Yang J.Y., Crowther J.A., Lilley P.E., Carr P.D., Ollis D.L., Brown S.E., Dixon N.E. Contrary to a classic phosphatase assay, the phosphate group is not released; instead, the p-nitrophenyl ester is cleaved off the of thymidine 5-monophosphate through hydrolysis. The release of p-nitrophenylate anions was measured spectrophotometrically. A divalent metal ion (e.g. Mg2+, Mn2+) is required for this reaction. All reactions were carried out at 25C and pH 8.00 except for those in Fig. 5 were the pH varied from 6.52 - 8.95. dixon@rsc.anu.edu.au Accepted 2013-JUN-26 by MGT 02-Aug-2013: Contacted by IntAct-Help. Biochemistry (0006-2960) 2002 Only protein-protein interactions imex curation Hydrolysis of the 5'-p-nitrophenyl ester of TMP by the proofreading exonuclease (epsilon) subunit of Escherichia coli DNA polymerase III. Hydrolysis of the 5'-p-nitrophenyl ester of TMP by the proofreading exonuclease (epsilon) subunit of Escherichia coli DNA polymerase III. Biochemistry (0006-2960) 2002 imex curation Hamdan S., Bulloch E.M., Thompson P.R., Beck J.L., Yang J.Y., Crowther J.A., Lilley P.E., Carr P.D., Ollis D.L., Brown S.E., Dixon N.E. dixon@rsc.anu.edu.au 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro ion exchange chrom ion exchange chromatography IEC IEC ion exchange chrom weight by staining molecular weight estimation by staining weight by staining Hamdan S., Bulloch E.M., Thompson P.R., Beck J.L., Yang J.Y., Crowther J.A., Lilley P.E., Carr P.D., Ollis D.L., Brown S.E., Dixon N.E. dixon@rsc.anu.edu.au Accepted 2013-JUN-26 by MGT 02-Aug-2013: Contacted by IntAct-Help. Biochemistry (0006-2960) 2002 Only protein-protein interactions imex curation hole_ecoli DNA polymerase III subunit theta holE b1842 JW1831 protein protein ecoli Escherichia coli (strain K12) MLKNLAKLDQTEMDKVNVDLAAAGVAFKERYNMPVIAEAVEREQPEHLRSWFRERLIAHRLASVNLSRLPYEPKLK 3667D87327E96556 3667D87327E96556 tmp thymidine 5'-monophosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pnp-tmp p-nitrophenyl ester of thymidine 5'-monophosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dpo3e_ecoli DNA polymerase III subunit epsilon dnaQ mutD b0215 JW0205 protein protein ecoli Escherichia coli (strain K12) MSTAITRQIVLDTETTGMNQIGAHYEGHKIIEIGAVEVVNRRLTGNNFHVYLKPDRLVDPEAFGVHGIADEFLLDKPTFAEVADEFMDYIRGAELVIHNAAFDIGFMDYEFSLLKRDIPKTNTFCKVTDSLAVARKMFPGKRNSLDALCARYEIDNSKRTLHGALLDAQILAEVYLAMTGGQTSMAFAMEGETQQQQGEATIQRIVRQASKLRVVFATDEEIAAHEARLDLVQKKGGSCLWRA DIP protein P03007 original sequence version: 106 DIP protein P03007 original sequence version: null 4211C9A00744964F dnaq-hole-1 2 3 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. S3b, S4b dnaq-hole-2 1 3 cofactor cofactor coenzyme neutral component neutral component ecoli Escherichia coli (strain K12) 6 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage Fig.1b dnaq-pnp_tmp-1 1 5 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 6 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) cleavage cleavage reaction cleavage Fig.1, 3, 4,5 Km and Kcat values are taken from Figure 1. dnaq-pnp_tmp-2 1 5 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 6 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 4 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage Fig. 4