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 NleG Type 3 effectors from enterohaemorrhagic Escherichia coli are U-Box E3 ubiquitin ligases. NleG Type 3 effectors from enterohaemorrhagic Escherichia coli are U-Box E3 ubiquitin ligases. PLoS pathogens 2010 imex curation Wu B., Skarina T., Yee A., Jobin MC., Dileo R., Semesi A., Fares C., Lemak A., Coombes BK., Arrowsmith CH., Singer AU., Savchenko A. alexei.savchenko@utoronto.ca 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Wu B., Skarina T., Yee A., Jobin MC., Dileo R., Semesi A., Fares C., Lemak A., Coombes BK., Arrowsmith CH., Singer AU., Savchenko A. alexei.savchenko@utoronto.ca Accepted 2020-MAR-04 AT 15:57 GMT AT 15:57 GMT by LUANA PLoS pathogens 2010 Only protein-protein interactions imex curation imex curation NleG Type 3 effectors from enterohaemorrhagic Escherichia coli are U-Box E3 ubiquitin ligases. NleG Type 3 effectors from enterohaemorrhagic Escherichia coli are U-Box E3 ubiquitin ligases. PLoS pathogens 2010 imex curation Wu B., Skarina T., Yee A., Jobin MC., Dileo R., Semesi A., Fares C., Lemak A., Coombes BK., Arrowsmith CH., Singer AU., Savchenko A. alexei.savchenko@utoronto.ca 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Wu B., Skarina T., Yee A., Jobin MC., Dileo R., Semesi A., Fares C., Lemak A., Coombes BK., Arrowsmith CH., Singer AU., Savchenko A. alexei.savchenko@utoronto.ca Accepted 2020-MAR-04 AT 15:57 GMT AT 15:57 GMT by LUANA PLoS pathogens 2010 Only protein-protein interactions imex curation imex curation ub2d2_human Ubiquitin-conjugating enzyme E2 D2 UBC5B UBCH4 UBE2D2 UBC4 UBCH5B Ubiquitin-protein ligase D2 Ubiquitin carrier protein D2 Ubiquitin-conjugating enzyme E2-17 kDa 2 Ubiquitin-conjugating enzyme E2(17)KB 2 PUBC1 p53-regulated ubiquitin-conjugating enzyme 1 (E3-independent) E2 ubiquitin-conjugating enzyme D2 E2 ubiquitin-conjugating enzyme D2 protein protein human Homo sapiens Human MALKRIHKELNDLARDPPAQCSAGPVGDDMFHWQATIMGPNDSPYQGGVFFLTIHFPTDYPFKPPKVAFTTRIYHPNINSNGSICLDILRSQWSPALTISKVLLSICSLLCDPNPDDPLVPEIARIYKTDREKYNRIAREWTQKYAM DIP protein P62837 original sequence version: 81 DIP protein P62837-1 original sequence version: 134 DIP protein P62837 original sequence version: 95 DIP protein P62837 original sequence version: 139 DIP protein P62837 original sequence version: 37 DIP protein P62837 original sequence version: 115 DIP protein P62837 original sequence version: 58 DIP protein P62837 original sequence version: 74 DIP protein P62837 original sequence version: 103 DIP protein P62837 original sequence version: 101 C942BE7853CBC355 q8x4x3_eco57 nleG5-1 ECs_1996 E5F08_13970 E5F08_18645 protein protein eco57 Escherichia coli O157:H7 MPVDLTPYILPGVSFLSDIPQETLSEIRNQTIRGEAQIRLGELMVSIRPMQVNGYFMGSLNQDGLSNDNIQIGLQYIEHIERTLNHGSLTSREVTVLREIEMLENMDLLSNYQLEELLDKIEVCAFNVEHAQLQVPESLRTCPVTLCEPEDGVFMRNSMNSNVCMLYDKMALIHLVKTRAAHPLSRESIAVSMIVGRDNCAFDPDRGNFVLKN 0AA796D5D4DBCA32 q8x509_eco57 nleG2-3 ECs_2156 E5F08_13145 protein protein eco57 Escherichia coli O157:H7 MPLTSDIRSHSFNLGVEVVRARIVANGRGDITVGGETVSIVYDSTNGRFSSSGGNGGLLSELLLLGFNSGPRALGERMLSMLSDSGEAQSQESIQNKISQCKFSVCPERLQCPLEAIQCPITLEQPEKGIFVKNSDGSDVCTLFDAAAFSRLVGEGLPHPLTREPITASIIVKHEECIYDDTRGNFIIKGN 8AF8F72983C88328 nleg2_3-ube2d2-1 2 3 unspecified role unspecified role neutral component neutral component purified purified ecolx Escherichia coli 5 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction fig. 6a-b nleg5_1-ube2d2-1 1 4 unspecified role unspecified role neutral component neutral component purified purified ecolx Escherichia coli 3 unspecified role unspecified role neutral component neutral component purified purified P62837:p.Phe62Ala mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain F A Thu Mar 05 22:52:53 GMT 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.phe62ala' Tue Jun 30 19:49:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe62Ala' tag fluor-5-maleimide fluorescein-5-maleimide label fluor-5-maleimide undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P62837:p.Arg5Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A Thu Mar 05 22:16:14 GMT 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.arg5ala' Tue Jun 30 19:35:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg5Ala' P62837:p.Lys63Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A Thu Mar 05 22:39:01 GMT 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.lys63ala' Tue Jun 30 17:02:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys63Ala' ecolx Escherichia coli direct interaction direct interaction fig. 7c To quantify the NleG5-1[1–213] interactions with UBE2D2 variants, we labelled the three UBE2D2 mutant proteins described above with fluorescein-5-maleimide and monitored the change in fluorescence polarization upon titration of NleG5-1[1–213] (Figure 7C). Kd values for NleG5-1[1–213] interactions with UBE2D2 R5A, F62A and K63A mutants were 326±35, 36±2 and 213±9 µM, respectively. Compared to the Kd for NleG5-1[1–213] interaction with wild type UBE2D2, presented above (56±2 µM) the affinity of R5A and K63A mutants were approximately 5 and 3 times lower, respectively, while the affinity of the F62A mutant was slightly higher.