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 Lens epithelium-derived growth factor deSumoylation by Sumo-specific protease-1 regulates its transcriptional activation of small heat shock protein and the cellular response. Lens epithelium-derived growth factor deSumoylation by Sumo-specific protease-1 regulates its transcriptional activation of small heat shock protein and the cellular response. FEBS J. (1742-464X) 2012 Ishihara K., Fatma N., Bhargavan B., Chhunchha B., Kubo E., Dey S., Takamura Y., Kumar A., Singh DP. human-lec Homo sapiens lec lec anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ishihara K., Fatma N., Bhargavan B., Chhunchha B., Kubo E., Dey S., Takamura Y., Kumar A., Singh DP. FEBS J. (1742-464X) 2012 Lens epithelium-derived growth factor deSumoylation by Sumo-specific protease-1 regulates its transcriptional activation of small heat shock protein and the cellular response. Lens epithelium-derived growth factor deSumoylation by Sumo-specific protease-1 regulates its transcriptional activation of small heat shock protein and the cellular response. FEBS J. (1742-464X) 2012 Ishihara K., Fatma N., Bhargavan B., Chhunchha B., Kubo E., Dey S., Takamura Y., Kumar A., Singh DP. human-lec Homo sapiens lec lec fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Ishihara K., Fatma N., Bhargavan B., Chhunchha B., Kubo E., Dey S., Takamura Y., Kumar A., Singh DP. FEBS J. (1742-464X) 2012 sumo1_human Small ubiquitin-related modifier 1 SUMO1 Sentrin Ubiquitin-like protein SMT3C Ubiquitin-homology domain protein PIC1 Ubiquitin-like protein UBL1 GAP-modifying protein 1 SMT3 homolog 3 SMT3C SMT3H3 UBL1 OK/SW-cl.43 protein protein human Homo sapiens Human MSDQEAKPSTEDLGDKKEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNSLRFLFEGQRIADNHTPKELGMEEEDVIEVYQEQTGGHSTV DIP protein P63165 original sequence version: 36 DIP protein P63165 original sequence version: 40 DIP protein P63165 original sequence version: 124 DIP protein P63165 original sequence version: 125 DIP protein P63165 original sequence version: 83 DIP protein P63165 original sequence version: 85 DIP protein P63165 original sequence version: 81 DIP protein P63165 original sequence version: 84 DIP protein P63165 original sequence version: 103 DIP protein P63165 original sequence version: 100 DIP protein P63165 original sequence version: 95 DIP protein P63165 original sequence version: 98 DIP protein P63165 original sequence version: 101 DIP protein P63165-1 original sequence version: 131 DIP protein P63165 original sequence version: 102 DIP protein P63165 original sequence version: 122 DIP protein P63165 original sequence version: 62 DIP protein P63165 original sequence version: 119 DIP protein P63165 original sequence version: 82 89BE97D2D054FB33 psip1_human PC4 and SFRS1-interacting protein PSIP1 DFS70 LEDGF PSIP2 Lens epithelium-derived growth factor Transcriptional coactivator p75/p52 Dense fine speckles 70 kDa protein CLL-associated antigen KW-7 protein protein human Homo sapiens Human MTRDFKPGDLIFAKMKGYPHWPARVDEVPDGAVKPPTNKLPIFFFGTHETAFLGPKDIFPYSENKEKYGKPNKRKGFNEGLWEIDNNPKVKFSSQQAATKQSNASSDVEVEEKETSVSKEDTDHEEKASNEDVTKAVDITTPKAARRGRKRKAEKQVETEEAGVVTTATASVNLKVSPKRGRPAATEVKIPKPRGRPKMVKQPCPSESDIITEEDKSKKKGQEEKQPKKQPKKDEEGQKEEDKPRKEPDKKEGKKEVESKRKNLAKTGVTSTSDSEEEGDDQEGEKKRKGGRNFQTAHRRNMLKGQHEKEAADRKRKQEEQMETEQQNKDEGKKPEVKKVEKKRETSMDSRLQRIHAEIKNSLKIDNLDVNRCIEALDELASLQVTMQQAQKHTEMITTLKKIRRFKVSQVIMEKSTMLYNKFKNMFLVGEGDSVITQVLNKSLAEQRQHEEANKTKDQGKKGPNKKLEKEQTGSKTLNGGSDAQDGNQPQHNGESNEDSKDNHEASTKKKPSSEERETEISLKDSTLDN DIP protein O75475 original sequence version: 124 DIP protein O75475-1 original sequence version: 108 DIP protein O75475-1 original sequence version: 113 DIP protein O75475 original sequence version: 113 4B653D02B1D0E174 psip1-sumo1 1 SUMO1 3 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined LEDGF 4 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level association association F1D homomint mint Results revealed the presence of three bands with anti-LEDGF polyclonal antibody (Fig. 1D; a, lane 2: unSumoylated endogenous LEDGF, ~75kDa; endogenous Sumoylated LEDGF, ~87kDa and endogenous LEDGF Sumoylated by exogenous EGFP-Sumo1, ~115kDa). The same membrane was probed with anti- Sumo1 that gave rise two bands (Fig. 1D; a, lane 4: endogenously Sumoylated LEDGF, ~87kDa and endogenous LEDGF Sumoylated with exogenous EGFP-Sumo1, ~115kDa), while only one band could be detected when the same membrane probed with anti-GFP antibody (Fig. 1D; a, lane 6: endogenous LEDGF Sumoylated with EGFP-Sumo1). sumo1-psip1 1 SUMO1 3 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined LEDGF 4 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level association association F1E homomint mint To validate further Sumoylation of endogenous LEDGF, nuclear extracts were immunoprcipitated with anti-Sumo1 antibody and immunoblotted with anti-LEDGF antibody (Fig. 1E; lane 1) or anti-Sumo1 antibody. Cells transfected with EGFP-empty vector were taken as control (lanes 1 and 3). The data revealed the presence of two bands; Sumoylated LEDGF (~87kDa) and endogenous LEDGF Sumoylated with exogenous EGFP-Sumo1 (~115kDa). psip1-sumo1-1 1 LEDGF 4 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined SUMO1 3 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association F5A F5B homomint mint wild type EGFP-LEDGF or endogenous LEDGF was Sumoylated, and therefore migrated more slowly on gel, yielding band: EGFP-LEDGF plus EGFPSumo1 (Fig. 5A, ~143kDa, lane 2), endogenous LEDGF plus EGFP-Sumo1 (Fig. 5A, ~115kDa, lane 1, 2, 3), endogenous LEDGF plus endogenous Sumo1 (~87kDa, lane 1, 2, 3). Next, the same blotted membrane was probed with anti-LEDGF (Fig. 5 B) or anti-GFP (Fig.5 C) psip1-sumo1-2 2 SUMO1 3 tag fluorescence tag visualisation by fluorescence tag fluorescence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined LEDGF 4 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization F1c homomint mint