l aspartic acid β hydroxamate  (MedChemExpress)

Journal: Cell Death & Disease

Article Title: SENP3 promotes renal tubular epithelial cell apoptosis after ischemia-reperfusion injury via ASS1 deSUMOylation

doi: 10.1038/s41419-025-08308-2

Figure Lengend Snippet: a , b The interaction between SUMO2/3 and ASS1 was reduced in IRI kidney tissue and H/R-treated TCMK1 cells. c , d Endogenous binding between SENP3 and ASS1 in TCMK1 cells was enhanced following H/R. e , f SUMO2/3 conjugation occurred preferentially at lysine residues K239 and K310 of ASS1, as the double mutant ASS1-2KR(K239R/K310R) abrogated this modification. g The ASS1-2KR mutation impaired the interaction between SENP3 and ASS1. For co-immunoprecipitation assays in ( c – g ), HEK-293T cells were transfected for 48 h with expression vectors encoding ASS1, UBC9 (an essential E2-conjugating enzyme in the SUMOylation process), and SUMO2/3. All experiments were repeated in at least three biological replicates.

Article Snippet: Pharmacological inhibition was performed with the ASS1 inhibitor α-Methyl-DL-aspartic acid (MDLA, 20 mM, 24 h; MCE, HY-W142119).

Techniques: Binding Assay, Conjugation Assay, Mutagenesis, Modification, Immunoprecipitation, Transfection, Expressing

Journal: Cell Death & Disease

Article Title: SENP3 promotes renal tubular epithelial cell apoptosis after ischemia-reperfusion injury via ASS1 deSUMOylation

doi: 10.1038/s41419-025-08308-2

Figure Lengend Snippet: a , b Following H/R treatment, TCMK1 cells showed increased nuclear accumulation of ASS1, which was predominantly in a deSUMOylation form, as demonstrated by co-immunoprecipitation of nuclear fractions. c , d Nuclear ASS1 levels in TCMK1 cells transfected with control (NC) or SENP3-targeting siRNA after H/R. e , f Nuclear ASS1 expression in TCMK1 cells transduced with lentivirus encoding wild-type SENP3 (SENP3-WT) or catalytically inactive SENP3-C526S after H/R. The SENP3-C526S mutant failed to promote ASS1 nuclear accumulation. g Immunofluorescence staining further confirmed that the nuclear translocation of ASS1 induced by H/R was dependent on SENP3 activity. Scale bar, 20 μm. Data: mean ± SEM (n = 3) . ns, not significance. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Pharmacological inhibition was performed with the ASS1 inhibitor α-Methyl-DL-aspartic acid (MDLA, 20 mM, 24 h; MCE, HY-W142119).

Techniques: Immunoprecipitation, Transfection, Control, Expressing, Transduction, Mutagenesis, Immunofluorescence, Staining, Translocation Assay, Activity Assay

Journal: Cell Death & Disease

Article Title: SENP3 promotes renal tubular epithelial cell apoptosis after ischemia-reperfusion injury via ASS1 deSUMOylation

doi: 10.1038/s41419-025-08308-2

Figure Lengend Snippet: a , b TCMK1 cells were transfected with wild-type ASS1 (ASS-WT) or the SUMOylation-deficient mutant (ASS1-2KR) for 24 h and then subjected to H/R. Flow cytometry analysis (Annexin V-FITC/PI) and Western blotting of apoptosis-related proteins showed that ASS1-WT promoted apoptosis more strongly than ASS1-2KR. c , d TCMK1 cells were co-transfected with SENP3 siRNA and either ASS1-WT or ASS1-2KR for 24 h, followed by H/R. Apoptosis analysis and Western blot of apoptotic markers revealed that ASS1-WT, but not ASS1-2KR, significantly enhanced apoptosis even under SENP3-deficient conditions. Data: mean ± SEM ( n = 3). ns, not significance. **** p < 0.0001.

Article Snippet: Pharmacological inhibition was performed with the ASS1 inhibitor α-Methyl-DL-aspartic acid (MDLA, 20 mM, 24 h; MCE, HY-W142119).

Techniques: Transfection, Mutagenesis, Flow Cytometry, Western Blot

Journal: Cell Death & Disease

Article Title: SENP3 promotes renal tubular epithelial cell apoptosis after ischemia-reperfusion injury via ASS1 deSUMOylation

doi: 10.1038/s41419-025-08308-2

Figure Lengend Snippet: a TCMK1 cells were pretreated with the ASS1 Inhibitor MDLA (20 mM, 24 h) prior to H/R. MDLA reduced the expression of apoptosis-related proteins following H/R. b – d ASS1 knockdown in TCMK1 cells attenuated H/R-induced apoptosis, as assessed by Annexin-V-FITC/PI flow cytometry, and decreased the expression of apoptosis-related proteins. e Knockdown of SENP3 attenuated Trp53 transcriptional activity in TCMK1 cells under H/R conditions. f ASS1 knockdown suppressed H/R-induced Trp53 transcriptional activation. g The catalytically inactive SENP3 mutant (C526S) failed to enhance Trp53 transcriptional activity following H/R. Data: mean ± SEM ( n = 3). ns, not significance. * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: Pharmacological inhibition was performed with the ASS1 inhibitor α-Methyl-DL-aspartic acid (MDLA, 20 mM, 24 h; MCE, HY-W142119).

Techniques: Expressing, Knockdown, Flow Cytometry, Activity Assay, Activation Assay, Mutagenesis

Journal: Cell Death & Disease

Article Title: SENP3 promotes renal tubular epithelial cell apoptosis after ischemia-reperfusion injury via ASS1 deSUMOylation

doi: 10.1038/s41419-025-08308-2

Figure Lengend Snippet: Renal ischemia/reperfusion promotes SENP3-mediated deSUMOylation of ASS1, facilitating its nuclear accumulation and enhances Trp53 transcriptional activity, ultimately resulting in tubular epithelial cell apoptosis.

Article Snippet: Pharmacological inhibition was performed with the ASS1 inhibitor α-Methyl-DL-aspartic acid (MDLA, 20 mM, 24 h; MCE, HY-W142119).

Techniques: Activity Assay