Discovery of novel inhibitors of human S-adenosylmethionine decarboxylase based on in silico high-throughput screening and a non-radioactive enzymatic assay

Liao, Chenzeng; Wang, Yanlin; Tan, Xiao; Sun, Lidan; Liu, Sen

doi:10.1038/srep10754

Cited by 21 publications

(35 citation statements)

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“…The expression and purification of human AdoMetDC were similar as mentioned before (Liao et al, 2015;Ai et al, 2016). Briefly, the coding sequence of AdoMetDC was inserted in pET15b, and the plasmid was transformed into the Escherichia coli strain BL21(DE3).…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

“…The AdoMetDC-PEPC-MDH assay was developed in our previous study (Liao et al, 2015) and used to evaluate the activity of AdoMetDC with the procedures similar as before (Liao et al, 2015;Ai et al, 2016). Briefly, AdoMetDC was mixed with DMSO, MGBG or compounds, respectively, in wells and incubated at 37 • C for 30 min before R2 [400 unit/L phosphoenolpyruvate carboxylase (PEPC), 600 unit/L malate dehydrogenase (MDH), 0.45 mM NADH] was added to the wells in a 96-well plate.…”

Section: Adometdc-pepc-mdh Assaymentioning

confidence: 99%

“…We used the AdoMetDC-PEPC-MDH assay developed in our previous study (Liao et al, 2015) to experimentally validate the inhibitory potency of the purchased compounds on AdoMetDC's enzymatic activity. This assay quantifies the generation of CO 2 from the decarboxylation reaction and is suitable for the initial semi-quantitative screening.…”

Section: Experimental Validation Of Potential Adometdc Inhibitorsmentioning

confidence: 99%

“…The production of dcAdoMet is generated from the decarboxylation of S-adenosylmethionine (AdoMet), which is catalyzed by AdoMetDC (AdoMet decarboxylase) ( Figure 1A). Therefore, AdoMetDC is a rate-limiting enzyme in the biosynthesis of polyamines (Liao et al, 2015). To inhibit polyamine synthesis, many AdoMetDC inhibitors have been developed, including the first-generation inhibitor methylglyoxal bis (guanylhydrazone) (MGBG), the second-generation inhibitor SAM486A (Sardomozide, also known as CGP48664), and the third-generation inhibitor AbeAdo (5 -([(Z)-4-amino-2butenyl]methylamino)-5 -deoxy-adenosine), etc.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, we described the computer-aided discovery of both non-covalent and covalent AdoMetDC inhibitors based on the in silico protein design strategy (Liao et al, 2015;Ai et al, 2016). To assist the discovery of covalent AdoMetDC inhibitors, we presented a docking strategy named as steric-clashes alleviating receptors (SCAR) (Ai et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Repurposing Clinical Drugs as AdoMetDC Inhibitors Using the SCAR Strategy

et al. 2020

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With the escalating costs in drug development, discovering new uses of approved drugs, i.e., drug repurposing, has attracted increasing interest. Spermidine and spermine are important polyamines for most cells and their biosynthesis are strictly regulated by the polyamine metabolic network. In cancerous cells and tumor environments, the concentrations of polyamines are much higher than in normal cells. During the synthesis of spermidine and spermine, an amino-propyl group is provided by decarboxylated S-adenosylmethionine, and the latter is generated from S-adenosylmethionine by AdoMetDC (AdoMet decarboxylase). Therefore, as a ratelimiting enzyme in the biosynthesis of spermidine and spermine, AdoMetDC has been an attractive drug target in cancer studies. In the last decades, many AdoMetDC inhibitors have been discovered, and several AdoMetDC inhibitors are under clinical trials, but unfortunately, none of them have been approved yet. To overcome the high costs in time and money for discovering de novo inhibitors, we set out to repurpose clinic drugs as AdoMetDC inhibitors. We used steric-clashes alleviating receptors (SCAR), a computer-aided drug discovery strategy developed by us recently for in silico screening. By combining computational screening and experimental validation, we successfully identified two approved drugs that have inhibitory potency on AdoMetDC's enzymatic activity. SCAR was previously shown to be suitable for the discovery of both covalent and non-covalent inhibitors, and this work further demonstrated the value of the SCAR strategy in drug repurposing.

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Section: Protein Expression and Purificationmentioning

confidence: 99%

Section: Adometdc-pepc-mdh Assaymentioning

confidence: 99%

Section: Experimental Validation Of Potential Adometdc Inhibitorsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Repurposing Clinical Drugs as AdoMetDC Inhibitors Using the SCAR Strategy

et al. 2020

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Subtype‐specific transcription factors affect polyamine metabolism and the tumor microenvironment in breast cancer

Song,

Wang,

Liu

2024

Cancer Innovation

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BackgroundPolyamines play important roles in cell growth and proliferation. Polyamine metabolism genes are dysregulated in various tumors. Some polyamine metabolism genes are regulated by transcription factors. However, the transcription factors that regulate polyamine metabolism genes have not been completely identified. Additionally, whether any of the transcriptional regulations depend on tumor heterogeneity and the tumor microenvironment has not been investigated.MethodsWe used bulk RNA‐seq data to identify dysregulated polyamine metabolism genes and their transcription factors across breast cancer subtypes. Genes highly correlated with polyamine changes were obtained, and their subtype‐specific expressions were checked in tumor microenvironment cells using single‐cell RNA (scRNA)‐seq data. Gene Ontology enrichment analysis was used to explore their molecular functions and biological processes, and survival analysis was used to examine the impact of these genes on therapeutic outcome.ResultsWe first analyzed the dysregulation of polyamine synthesis, catabolism, and transport in four breast cancer subtypes. Genes such as AGMAT and CAV1 were dysregulated across all subtypes, while APRT, SAT1, and other genes were dysregulated in the more lethal subtypes. Among the dysregulated genes of polyamine metabolism, we focused on three genes (SRM, APRT, and SAT1) and identified their transcription factors (SPI1 and IRF1 correspond to SAT1, and IRF3 corresponds to SRM and APRT). With scRNA‐seq data, we verified that these three transcription factors also regulated these three polyamine metabolism genes in the tumor microenvironment. Both bulk RNA‐seq and scRNA‐seq data indicated that these genes were specifically upregulated in high‐risk breast cancer subtypes, such as the basal‐like type. High expression of these genes corresponded to worse outcomes in the basal‐like subtype under chemotherapy and radiation treatment.ConclusionOur work identified three subtype‐specific transcription factors that regulate three polyamine metabolism genes in high‐risk breast cancer subtypes and the tumor microenvironment. Our results deepen the understanding of the role of polyamine metabolism in breast cancer and may help the clinical therapy of advanced breast cancer subtypes.

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Supplementation of arginine, ornithine and citrulline in rainbow trout (Oncorhynchus mykiss): Effects on growth, amino acid levels in plasma and gene expression responses in liver tissue

Clark

Tinsley

Sigholt

et al. 2020

Comparative Biochemistry and Physiology Part A: Molecular &

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Functional amino acids (FAA) regulate metabolic pathways directly linked to health, survival, growth and development. Arginine is a FAA with crucial roles in protein deposition and the immune response.In mammals, supplementation of arginine's precursor amino acid, citrulline, is known to increase circulating arginine to levels beyond direct arginine supplementation, however, citrulline supplementation is poorly studied in fish. To address this knowledge gap, we supplemented the diet of rainbow trout with arginine and its precursor amino acids, ornithine and citrulline, at 3 levels (0.5%, 1% and 2% of the total diet) during a 14-week experiment. We sampled fish at 3h and 24h post-feeding to investigate immediate and steady-state effects, respectively. There were no differences in fish growth for any of the diets across a range of indicators. In blood plasma, out of 26 amino acids detected, 11 and 6 displayed significant changes 24h and 3h post-prandial, respectively. Arginine, ornithine and citrulline levels were all significantly increased by the citrulline supplemented diets. In muscle, 8 amino acids were significantly altered by supplemented diets, while there were no significant changes in liver.Arginine was increased by 2% citrulline supplementation in muscle tissue. We also investigated the transcriptional responses of urea cycle, nitric oxide cycle and rate-limiting polyamine synthesis enzymes, related to arginine's metabolism, in liver. At both time points, only 2 enzymes were significantly altered by the supplemented diets, however several significant changes were observed comparing 3h and 24h post-prandial expression levels. Of these, the paralogous polyamine synthesis enzyme encoding genes ODC1 and ODC2 displayed the largest increases in 3h post-prandial fish. These findings demonstrate that endogenous synthesis of arginine is possible from a citrulline supplemented diet and improve our understanding of arginine metabolism in fish.

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Discovery of novel inhibitors of human S-adenosylmethionine decarboxylase based on in silico high-throughput screening and a non-radioactive enzymatic assay

Cited by 21 publications

References 56 publications

Repurposing Clinical Drugs as AdoMetDC Inhibitors Using the SCAR Strategy

Repurposing Clinical Drugs as AdoMetDC Inhibitors Using the SCAR Strategy

Subtype‐specific transcription factors affect polyamine metabolism and the tumor microenvironment in breast cancer

Supplementation of arginine, ornithine and citrulline in rainbow trout (Oncorhynchus mykiss): Effects on growth, amino acid levels in plasma and gene expression responses in liver tissue

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