recombinant parp 1  (Novus Biologicals)

Journal: Advanced Science

Article Title: Novel Cyano‐Artemisinin Dimer ZQJ29 Targets PARP1 to Induce Ferroptosis in Pancreatic Cancer Treatment

doi: 10.1002/advs.202501935

Figure Lengend Snippet: ZQJ29 inhibits PARP1 activity through direct binding with PARP1. A) PANC‐1 and KP4 cells were treated with varying concentrations of ZQJ29 for 24 h. B,C) Statistical analysis of Figure . D) PANC‐1 and KP4 cells were treated with 5 µM of ZQJ29 for specified periods. E,F) Statistical analysis of Figure . G) Molecular docking model illustrating the binding interaction between ZQJ29 and PARP1. H) Immunofluorescence staining of PARP1 (red) and nuclear DAPI staining (blue) in PANC‐1 and KP4 cells after 24 h treatment with ZQJ29 (0, 1, 2.5, and 5 µ m ). Scale bar: 200 µm. I) Thermal stability analysis of PARP1‐ZQJ29 interaction using CETSA across a temperature gradient (45‐70 °C). (J–K) Statistical analysis of Figure . L) PARP1 stability at 60 °C under treatment with different ZQJ29 concentrations. M,N) Statistical analysis of Figure . O) SPR assay. P) Stability of PARP1 treated with varying pronase/protein ratios. Q,R) Statistical analysis of Figure . S) PARP1 stability under different ZQJ29 concentrations (1:3000). T,U) Statistical analysis of Figure . The data was shown as mean value ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant.

Article Snippet: Recombinant human PARP1 protein (Sino Biological, 11040‐H08B) was immobilized on an activated carboxymethylated 5 (CM5) sensor chip using the amine coupling method.

Techniques: Activity Assay, Binding Assay, Immunofluorescence, Staining, SPR Assay

Journal: Advanced Science

Article Title: Novel Cyano‐Artemisinin Dimer ZQJ29 Targets PARP1 to Induce Ferroptosis in Pancreatic Cancer Treatment

doi: 10.1002/advs.202501935

Figure Lengend Snippet: ZQJ29‐induced ferroptosis is PARP1‐dependent. A) PPI network analysis. B) Protein expression in mouse tumor tissues. C) Statistical analysis of Figure . D) Heatmap of protein expression from proteomic data. E) Protein expression in PANC‐1 and KP4 cells treated with varying concentrations of ZQJ29. F,G) Statistical analysis of Figure . H–J) Protein expression in PANC‐1 and KP4 cells treated with ZQJ29 alone or in combination with PARP1 inhibitor (Olaparib), SLC7A11 inhibitor (Erastin), or GPX4 inhibitor (ML‐210). K–P) Statistical analysis of Figure . Q) Expression of PARP1, TP53, SLC7A11, and GPX4 in PANC‐1 and KP4 cells transfected with different PARP1 siRNAs. R,S) Statistical analysis of Figure Q. T,U) Cell viability assessed by CCK‐8 assay in PANC‐1 and KP4 cells transfected with control‐siRNA or PARP1‐siRNA, followed by treatment with ZQJ29 (1 µ m ) for 24 h. The data was shown as mean value ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant.

Article Snippet: Recombinant human PARP1 protein (Sino Biological, 11040‐H08B) was immobilized on an activated carboxymethylated 5 (CM5) sensor chip using the amine coupling method.

Techniques: Expressing, Transfection, CCK-8 Assay, Control

Journal: Advanced Science

Article Title: Novel Cyano‐Artemisinin Dimer ZQJ29 Targets PARP1 to Induce Ferroptosis in Pancreatic Cancer Treatment

doi: 10.1002/advs.202501935

Figure Lengend Snippet: The schematic illustration for ZQJ29 targeted PARP1 to activate ferroptosis for anti‐pancreatic cancer.

Article Snippet: Recombinant human PARP1 protein (Sino Biological, 11040‐H08B) was immobilized on an activated carboxymethylated 5 (CM5) sensor chip using the amine coupling method.

Techniques:

Journal: Advanced Science

Article Title: Nudix Hydrolase 13 Impairs the Initiation of Colorectal Cancer by Inhibiting PKM1 ADP‐Ribosylation

doi: 10.1002/advs.202410058

Figure Lengend Snippet: PARP1‐induced PKM1 ADP‐ribosylation signals for ubiquitination and degradation. A) In vivo PARylation assay was performed in SW480 cells transfected with PKM1‐Myc. B) Control and NUDT13‐overexpressing SW480 cells pre‐treated with DMSO or Olaparib (10 µ m ) were exposed to 50µg mL −1 CHX for the indicated time. Quantification of PKM1 by densitometry. C) Immunoblot analysis of PKM1 ubiquitination levels in NUDT13‐overexpressing SW480 cells transfected with NUDT13 siRNA, and treated with or without Olaparib (10 µ m ) for 48h. (D and E) The interaction between PKM1 and PARP1 was detected by Co‐IP D) and in vitro pull‐down assays E). F) In vitro PARylation assay to detect the PARylation of purified PKM1 catalyzed by recombinant human PARP1. G) Immunoblot analysis of PKM1 PARylation levels in SW480 cells treated with Olaparib (10 µ m ) for 48h, or transfected with NUDT13‐Flag plasmids. H) Immunoblot analysis of PKM1 PARylation levels in 293T cells transfected with WT or mutant PKM1. The amounts of PKM1 levels in different groups were adjusted by MG132 treatment. I) In vitro PARylation assay to detect the PARylation of purified PKM1. J) CHX chase assays were conducted to assess the stabilities of PKM1 WT and A4 mutant. All results mentioned above were obtained from 3 or more independent experiments. Data are presented as mean ± SD; P value was calculated by two‐way ANOVA (B and J).

Article Snippet: The purified recombinant PKM1, H1, H2A, or H2B (1 µg) was mixed with 1 µg recombinant human PARP1 (Sino Biological) with or without NAD + (HY‐B0445, MCE, China) in 1×reaction buffer (50 m m Tris‐HCl (pH 8.0), 4 m m MgCl 2 , 2 0m m NaCl, 1 m m DTT, and 100 ng sheared DNA (D7656, Sigma–Aldrich, USA)) at 37 °C for 30 min.

Techniques: In Vivo, Transfection, Control, Western Blot, Co-Immunoprecipitation Assay, In Vitro, Purification, Recombinant, Mutagenesis