InhibitorProline modulationOncologyTool compoundFragment-based drug discoveryPyrroline-5-carboxylate reductase 1 (PYCR1) is the final enzyme involved in the biosynthesis of proline and has been found to be upregulated in various forms of cancer. Due to the role of proline in maintaining the redox...
Inhibiting PYCR1 genetically with small interfering RNA (siRNA)/short hairpin RNA (shRNA), or pharmacologically with a small molecule inhibitor (PYCR1i),35 was sufficient to decrease 13C5-proline synthesis from 13C5-glutamine (Fig. 3a–e and Extended Data Fig. 3a–e), while not substantially...
Western blot The cells were first washed with cold PBS three times, then lysed in RIPA lysis buffer (Beyotime, Shanghai, China) con- taining a complete protease inhibitor cocktail (Boehringer Mannheim, USA), and cell lysates were collected. Con- centrations of proteins were then determined by...
Cells were rinsed with PBS and lysed with buffer A (50 mM HEPES, pH 7.5; 70 mM potassium acetate and 5 mM magnesium acetate) supplemented with protease inhibitor cocktail (Sigma-Aldrich) and phosphatase inhibitors (10 mM β-glycerophosphate, 1 mM sodium fluoride and 0.1 mM sod...
Cells were rinsed with PBS and lysed with buffer A (50 mM HEPES, pH 7.5; 70 mM potassium acetate and 5 mM magnesium acetate) supplemented with protease inhibitor cocktail (Sigma-Aldrich) and phosphatase inhibitors (10 mM β-glycerophosphate, 1 mM sodium fluoride and 0.1 mM sod...
Each dish was added 2 mL cold PBS containing Protease Inhibitor Cocktail. The cells were collected with 800 g at 4ºC for 5 minutes. During spin, each sample was combined with 0.5 mL of cell Lysis Buffer with 2.5 µL of Protease Inhibitor Cocktail II. Tumor formation assay in nude ...
To further confirm this, we used IKK inhibitor to treat Lon-overexpressing cells regarding the activation of p38α signaling. We found that the Lon-induced EMT, but not the activation of p38α signaling, was inhibited by treatment with IKK inhibitor VII (Fig. 3C). Consistently, we found ...
The structure of PYCR1 complexed with NFLP shows that inhibitor binding is accompanied by conformational changes in the active site, including the translation of an a-helix by 1 . These changes are unique to NFLP and enable additional hydrogen bonds with the enzyme. NFLP was also shown to ...
Co-exposure of High-fat-diet (HFD) behavior and environmental low-dose radiation (LDR) is common among majority occupational workers, but the synergism of this co-exposure in metabolic health is poorly understood. This study aimed to investigate the impact of gut microbiota and its metabolites on...
Immunoprecipitation. Cells were rinsed with PBS and lysed with buffer A (50 mM HEPES, pH 7.5; 70 mM potassium acetate and 5 mM magnesium acetate) supplemented with protease inhibitor cocktail (Sigma-Aldrich) and phosphatase inhibitors (10 mM β -glycerophosphate, 1 mM sodium ...