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 Regulation of G6PD acetylation by SIRT2 and KAT9 modulates NADPH homeostasis and cell survival during oxidative stress. Regulation of G6PD acetylation by SIRT2 and KAT9 modulates NADPH homeostasis and cell survival during oxidative stress. EMBO J. (0261-4189) 2014 imex curation Wang Y.P., Zhou L.S., Zhao Y.Z., Wang S.W., Chen L.L., Liu L.X., Ling Z.Q., Hu F.J., Sun Y.P., Zhang J.Y., Yang C., Yang Y., Xiong Y., Guan K.L., Ye D. yedan@fudan.edu.cn 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang Y.P., Zhou L.S., Zhao Y.Z., Wang S.W., Chen L.L., Liu L.X., Ling Z.Q., Hu F.J., Sun Y.P., Zhang J.Y., Yang C., Yang Y., Xiong Y., Guan K.L., Ye D. yedan@fudan.edu.cn Accepted 2014-DEC-03 by MARTA EMBO J. (0261-4189) 2014 Only protein-protein interactions imex curation Regulation of G6PD acetylation by SIRT2 and KAT9 modulates NADPH homeostasis and cell survival during oxidative stress. Regulation of G6PD acetylation by SIRT2 and KAT9 modulates NADPH homeostasis and cell survival during oxidative stress. EMBO J. (0261-4189) 2014 imex curation Wang Y.P., Zhou L.S., Zhao Y.Z., Wang S.W., Chen L.L., Liu L.X., Ling Z.Q., Hu F.J., Sun Y.P., Zhang J.Y., Yang C., Yang Y., Xiong Y., Guan K.L., Ye D. yedan@fudan.edu.cn 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Wang Y.P., Zhou L.S., Zhao Y.Z., Wang S.W., Chen L.L., Liu L.X., Ling Z.Q., Hu F.J., Sun Y.P., Zhang J.Y., Yang C., Yang Y., Xiong Y., Guan K.L., Ye D. yedan@fudan.edu.cn Accepted 2014-DEC-03 by MARTA EMBO J. (0261-4189) 2014 Only protein-protein interactions imex curation g6pd_human Glucose-6-phosphate 1-dehydrogenase G6PD protein protein human Homo sapiens Human MAEQVALSRTQVCGILREELFQGDAFHQSDTHIFIIMGASGDLAKKKIYPTIWWLFRDGLLPENTFIVGYARSRLTVADIRKQSEPFFKATPEEKLKLEDFFARNSYVAGQYDDAASYQRLNSHMNALHLGSQANRLFYLALPPTVYEAVTKNIHESCMSQIGWNRIIVEKPFGRDLQSSDRLSNHISSLFREDQIYRIDHYLGKEMVQNLMVLRFANRIFGPIWNRDNIACVILTFKEPFGTEGRGGYFDEFGIIRDVMQNHLLQMLCLVAMEKPASTNSDDVRDEKVKVLKCISEVQANNVVLGQYVGNPDGEGEATKGYLDDPTVPRGSTTATFAAVVLYVENERWDGVPFILRCGKALNERKAEVRLQFHDVAGDIFHQQCKRNELVIRVQPNEAVYTKMMTKKPGMFFNPEESELDLTYGNRYKNVKLPDAYERLILDVFCGSQMHFVRSDELREAWRIFTPLLHQIELEKPKPIPYIYGSRGPTEADELMKRVGFQYEGTYKWVNPHKL F2B775340640A96F sir2_human NAD-dependent protein deacetylase sirtuin-2 SIR2-like protein 2 SIRT2 SIR2L SIR2L2 Regulatory protein SIR2 homolog 2 protein protein human Homo sapiens Human MAEPDPSHPLETQAGKVQEAQDSDSDSEGGAAGGEADMDFLRNLFSQTLSLGSQKERLLDELTLEGVARYMQSERCRRVICLVGAGISTSAGIPDFRSPSTGLYDNLEKYHLPYPEAIFEISYFKKHPEPFFALAKELYPGQFKPTICHYFMRLLKDKGLLLRCYTQNIDTLERIAGLEQEDLVEAHGTFYTSHCVSASCRHEYPLSWMKEKIFSEVTPKCEDCQSLVKPDIVFFGESLPARFFSCMQSDFLKVDLLLVMGTSLQVQPFASLISKAPLSTPRLLINKEKAGQSDPFLGMIMGLGGGMDFDSKKAYRDVAWLGECDQGCLALAELLGWKKELEDLVRREHASIDAQSGAGVPNPSTSASPKKSPPPAKDEARTTEREKPQ DIP protein Q8IXJ6 original sequence version: 113 A392442A8F6316F1 g6pd-g6pd-3 2 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western 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 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P11413:p.Lys403Gln mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K Q Wed May 02 17:48:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys403gln' Tue Jun 30 19:54:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys403Gln' physical association physical association F2B F2D SF5 F4E To test this hypothesis, we determined the interaction between two differentially tagged G6PD proteins, G6PD-Flag and GFP-G6PD, in HEK293T cells. We found that mutation of K171 to either R (K171R) or Q (K171Q) did not affect the interaction between G6PD subunits (Fig 2B). In contrast, substitution of K386Q, but not K386R, impaired the interaction between G6PD subunits (Fig 2B). Strikingly, substitution of K403Q, but not K403R, entirely disrupted the interaction between G6PD subunits (Fig 2B). In addition, glutaraldehyde cross-linking assay demonstrated that the K403Q mutant displayed impaired ability to form dimers when compared to wildtype G6PD or the K403R mutant (Fig 2C). Moreover, NAM treatment decreased the binding by approximately 55% between the two differentially tagged proteins of wild-type G6PD in a dose-dependent manner (Fig 2D and Supplementary Fig S5). NAM treatment, however, failed to affect the interaction between Flag-tagged and GFP-tagged K403R mutant of G6PD (Fig 2D), further suggesting that K403 acetylation largely hinders the interaction between G6PD subunits. Co-expression of SIRT2, but not the catalytic inactive SIRT2H187Y mutant, increased the interaction between two differentially tagged G6PD, and this effect could be diminished by NAM treatment (Fig 4E). g6pd-g6pd-4 2 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P11413:p.Lys386Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K Q Wed May 02 19:47:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys386gln' Tue Jun 30 17:23:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys386Gln' 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western 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 physical association physical association F2B To test this hypothesis, we determined the interaction between two differentially tagged G6PD proteins, G6PD-Flag and GFP-G6PD, in HEK293T cells. We found that mutation of K171 to either R (K171R) or Q (K171Q) did not affect the interaction between G6PD subunits (Fig 2B). In contrast, substitution of K386Q, but not K386R, impaired the interaction between G6PD subunits (Fig 2B). Strikingly, substitution of K403Q, but not K403R, entirely disrupted the interaction between G6PD subunits (Fig 2B). In addition, glutaraldehyde cross-linking assay demonstrated that the K403Q mutant displayed impaired ability to form dimers when compared to wildtype G6PD or the K403R mutant (Fig 2C). Moreover, NAM treatment decreased the binding by approximately 55% between the two differentially tagged proteins of wild-type G6PD in a dose-dependent manner (Fig 2D and Supplementary Fig S5). NAM treatment, however, failed to affect the interaction between Flag-tagged and GFP-tagged K403R mutant of G6PD (Fig 2D), further suggesting that K403 acetylation largely hinders the interaction between G6PD subunits. g6pd-g6pd-2 2 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f4a To test this hypothesis, we determined the interaction between two differentially tagged G6PD proteins, G6PD-Flag and GFP-G6PD, in HEK293T cells. We found that mutation of K171 to either R (K171R) or Q (K171Q) did not affect the interaction between G6PD subunits (Fig 2B). In contrast, substitution of K386Q, but not K386R, impaired the interaction between G6PD subunits (Fig 2B). Strikingly, substitution of K403Q, but not K403R, entirely disrupted the interaction between G6PD subunits (Fig 2B). In addition, glutaraldehyde cross-linking assay demonstrated that the K403Q mutant displayed impaired ability to form dimers when compared to wildtype G6PD or the K403R mutant (Fig 2C). Moreover, NAM treatment decreased the binding by approximately 55% between the two differentially tagged proteins of wild-type G6PD in a dose-dependent manner (Fig 2D and Supplementary Fig S5). NAM treatment, however, failed to affect the interaction between Flag-tagged and GFP-tagged K403R mutant of G6PD (Fig 2D), further suggesting that K403 acetylation largely hinders the interaction between G6PD subunits. sirt2-g6pd 2 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f4b To test this hypothesis, we determined the interaction between two differentially tagged G6PD proteins, G6PD-Flag and GFP-G6PD, in HEK293T cells. We found that mutation of K171 to either R (K171R) or Q (K171Q) did not affect the interaction between G6PD subunits (Fig 2B). In contrast, substitution of K386Q, but not K386R, impaired the interaction between G6PD subunits (Fig 2B). Strikingly, substitution of K403Q, but not K403R, entirely disrupted the interaction between G6PD subunits (Fig 2B). In addition, glutaraldehyde cross-linking assay demonstrated that the K403Q mutant displayed impaired ability to form dimers when compared to wildtype G6PD or the K403R mutant (Fig 2C). Moreover, NAM treatment decreased the binding by approximately 55% between the two differentially tagged proteins of wild-type G6PD in a dose-dependent manner (Fig 2D and Supplementary Fig S5). NAM treatment, however, failed to affect the interaction between Flag-tagged and GFP-tagged K403R mutant of G6PD (Fig 2D), further suggesting that K403 acetylation largely hinders the interaction between G6PD subunits. g6pd-sirt2 1 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association F6D To test this hypothesis, we determined the interaction between two differentially tagged G6PD proteins, G6PD-Flag and GFP-G6PD, in HEK293T cells. We found that mutation of K171 to either R (K171R) or Q (K171Q) did not affect the interaction between G6PD subunits (Fig 2B). In contrast, substitution of K386Q, but not K386R, impaired the interaction between G6PD subunits (Fig 2B). Strikingly, substitution of K403Q, but not K403R, entirely disrupted the interaction between G6PD subunits (Fig 2B). In addition, glutaraldehyde cross-linking assay demonstrated that the K403Q mutant displayed impaired ability to form dimers when compared to wildtype G6PD or the K403R mutant (Fig 2C). Moreover, NAM treatment decreased the binding by approximately 55% between the two differentially tagged proteins of wild-type G6PD in a dose-dependent manner (Fig 2D and Supplementary Fig S5). NAM treatment, however, failed to affect the interaction between Flag-tagged and GFP-tagged K403R mutant of G6PD (Fig 2D), further suggesting that K403 acetylation largely hinders the interaction between G6PD subunits.