ACS Bio & Med Chem Au最新文献

{"title":"Bacteriostatic and Immunomodulatory Effect of Xylocaine in the Context of <i>in Vitro</i> Bladder Epithelial Cell Infection.","authors":"John Kerr White, Yundi Yin, Soumitra Mohanty, Natalia Ferraz, Annelie Brauner","doi":"10.1021/acsbiomedchemau.4c00070","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00070","url":null,"abstract":"<p><p>The application of urinary catheters is associated with pain and discomfort. Several topical medications are available to ease catheter insertion, including xylocaine. Here we report that xylocaine, although not classified as an antibacterial agent, has bacteriostatic properties against both Gram-positive and Gram-negative etiological agents of urinary tract infections (UTIs). Xylocaine reduces the amount of biofilm formed by ESBL- and non-ESBL-producing <i>E. coli</i>. In addition, xylocaine possesses slight immunomodulatory properties in uroepithelial cells, and treatment of uroepithelial cells prior to infection reduces bacterial loads in the supernatant. In conclusion, xylocaine has multifaceted positive effects when used during the insertion of urinary catheters.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"31-34"},"PeriodicalIF":3.8,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843344/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bacteriostatic and Immunomodulatory Effect of Xylocaine in the Context of in Vitro Bladder Epithelial Cell Infection","authors":"John Kerr White,&nbsp;Yundi Yin,&nbsp;Soumitra Mohanty,&nbsp;Natalia Ferraz and Annelie Brauner*,&nbsp;","doi":"10.1021/acsbiomedchemau.4c0007010.1021/acsbiomedchemau.4c00070","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00070https://doi.org/10.1021/acsbiomedchemau.4c00070","url":null,"abstract":"<p >The application of urinary catheters is associated with pain and discomfort. Several topical medications are available to ease catheter insertion, including xylocaine. Here we report that xylocaine, although not classified as an antibacterial agent, has bacteriostatic properties against both Gram-positive and Gram-negative etiological agents of urinary tract infections (UTIs). Xylocaine reduces the amount of biofilm formed by ESBL- and non-ESBL-producing <i>E. coli</i>. In addition, xylocaine possesses slight immunomodulatory properties in uroepithelial cells, and treatment of uroepithelial cells prior to infection reduces bacterial loads in the supernatant. In conclusion, xylocaine has multifaceted positive effects when used during the insertion of urinary catheters.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"31–34 31–34"},"PeriodicalIF":3.8,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00070","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of Small Interfering RNA Loaded Cationic Lipid Nanoparticles for the Treatment of Liver Cancer with Elevated α-Fetoprotein Expression","authors":"Kongpop Duangchan,&nbsp;Nathachit Limjunyawong,&nbsp;Kamonlatth Rodponthukwaji,&nbsp;Teeranai Ittiudomrak,&nbsp;Mattika Thaweesuvannasak,&nbsp;Natsuda Kunwong,&nbsp;Chanatip Metheetrairut,&nbsp;Vorapan Sirivatanauksorn,&nbsp;Yongyut Sirivatanauksorn,&nbsp;Prawat Kositamongkol,&nbsp;Prawej Mahawithitwong,&nbsp;Chutwichai Tovikkai,&nbsp;Kytai T. Nguyen,&nbsp;Chatchawan Srisawat and Primana Punnakitikashem*,&nbsp;","doi":"10.1021/acsbiomedchemau.4c0006110.1021/acsbiomedchemau.4c00061","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00061https://doi.org/10.1021/acsbiomedchemau.4c00061","url":null,"abstract":"<p >α-Fetoprotein (AFP) is an oncogenic glycoprotein that is overexpressed in most patients with liver cancer. Moreover, it significantly affects tumorigenesis and progression, particularly by inhibiting programmed cell death or apoptosis. The treatment of liver cancer with chemotherapy is currently still in use, but its toxicity is a major concern. Alternatively, targeted therapy, especially small interfering RNA (siRNA)-based therapeutics that utilize siRNA to suppress target gene expression, is a promising cancer treatment approach that can help reduce such drawbacks. However, transporting siRNA into cells is a challenge due to its ease of degradation and limited cell membrane permeability. To overcome this limitation, we fabricated cationic lipid nanoparticles (cLNPs) to deliver <i>AFP-</i>targeted siRNA (siAFP) to AFP-producing liver cancer cells. Our results illustrated that these nanoparticles had a high capacity for siRNA encapsulation (&gt;95%) and entered the cancer cells efficiently. Cell internalization of siAFP-loaded cLNPs resulted in the silencing of <i>AFP</i> mRNA expression and led to increased apoptotic cell death by inducing caspase-3/7 activity. This suggested that our cLNPs could be used as a powerful siRNA delivery carrier and siAFP-loaded cLNPs might be a useful strategy for treating liver cancer in the future.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"78–88 78–88"},"PeriodicalIF":3.8,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of Small Interfering RNA Loaded Cationic Lipid Nanoparticles for the Treatment of Liver Cancer with Elevated α-Fetoprotein Expression.","authors":"Kongpop Duangchan, Nathachit Limjunyawong, Kamonlatth Rodponthukwaji, Teeranai Ittiudomrak, Mattika Thaweesuvannasak, Natsuda Kunwong, Chanatip Metheetrairut, Vorapan Sirivatanauksorn, Yongyut Sirivatanauksorn, Prawat Kositamongkol, Prawej Mahawithitwong, Chutwichai Tovikkai, Kytai T Nguyen, Chatchawan Srisawat, Primana Punnakitikashem","doi":"10.1021/acsbiomedchemau.4c00061","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00061","url":null,"abstract":"<p><p>α-Fetoprotein (AFP) is an oncogenic glycoprotein that is overexpressed in most patients with liver cancer. Moreover, it significantly affects tumorigenesis and progression, particularly by inhibiting programmed cell death or apoptosis. The treatment of liver cancer with chemotherapy is currently still in use, but its toxicity is a major concern. Alternatively, targeted therapy, especially small interfering RNA (siRNA)-based therapeutics that utilize siRNA to suppress target gene expression, is a promising cancer treatment approach that can help reduce such drawbacks. However, transporting siRNA into cells is a challenge due to its ease of degradation and limited cell membrane permeability. To overcome this limitation, we fabricated cationic lipid nanoparticles (cLNPs) to deliver <i>AFP-</i>targeted siRNA (siAFP) to AFP-producing liver cancer cells. Our results illustrated that these nanoparticles had a high capacity for siRNA encapsulation (>95%) and entered the cancer cells efficiently. Cell internalization of siAFP-loaded cLNPs resulted in the silencing of <i>AFP</i> mRNA expression and led to increased apoptotic cell death by inducing caspase-3/7 activity. This suggested that our cLNPs could be used as a powerful siRNA delivery carrier and siAFP-loaded cLNPs might be a useful strategy for treating liver cancer in the future.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"78-88"},"PeriodicalIF":3.8,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843345/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic Probing of Sialylated Glycoconjugates with Fluorine-Selenol Displacement Reaction (FSeDR).","authors":"Yue Zhao, Zhigang Lyu, Benjamin Prather, Todd R Lewis, Jinfeng Kang, Rongsheng E Wang","doi":"10.1021/acsbiomedchemau.4c00084","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00084","url":null,"abstract":"<p><p>Dysregulated sialic acid biosynthesis is characteristic of the onset and progression of human diseases including hormone-sensitive prostate cancer and breast cancer. The sialylated glycoconjugates involved in this process are therefore important targets for identification and functional studies. To date, one of the most common strategies is metabolic glycoengineering, which utilizes <i>N</i>-acetylmannosamine (ManNAc) analogues such as <i>N</i>-azidoacetylmannosamine (ManNAz) to hijack sialic acid biosynthesis and label the sialylated glycoconjugates with \"click chemistry (CuAAC)\" tags. Yet, current chemical modifications including those CuAAC-based alkyne/azide tags are still big in size, and the resulting steric hindrance perturbs the mannosamine and sialic acid derivatives' recognition and metabolism by enzymes involved in biosynthetic pathways. As a result, the peracetylated ManNAz has compromised incorporation to sialic acid substrates and manifests cellular growth inhibition and cytotoxicity. Herein, we show that the α-fluorinated peracetylated analogue ManN(F-Ac) displayed a satisfying safety profile in mammalian cell lines at concentrations as high as 500 μM. More importantly, aliphatic selenol-containing probes can efficiently displace α-fluorine in fluoroacetamide-containing substrates including ManN(F-Ac) at a neutral pH range (∼7.2). The combined use of peracetylated ManN(F-Ac) and the dethiobiotin-selenol probe as the fluorine-selenol displacement reaction (FSeDR) toolkit allowed for successful metabolic labeling of sialoglycoproteins in multiple prostate and cancer cell lines, including PC-3 and MDA-MB-231. More sialoglycoproteins in these cell lines were demonstrated to be labeled by FSeDR compared with the traditional CuAAC approach. Lastly, with FSeDR-mediated metabolic labeling, we were able to probe the cellular expression level and spatial distribution of sialylated glycoconjugates during the progression of these hormone-sensitive cancer cells. Taken together, the promising results suggest the potential of the FSeDR strategy to efficiently and systematically identify and study sialic acid substrates and potentially empower metabolic engineering on a diverse set of glycosylated proteins that are vital for human diseases.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"119-130"},"PeriodicalIF":3.8,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843343/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic Probing of Sialylated Glycoconjugates with Fluorine-Selenol Displacement Reaction (FSeDR)","authors":"Yue Zhao,&nbsp;Zhigang Lyu,&nbsp;Benjamin Prather,&nbsp;Todd R. Lewis,&nbsp;Jinfeng Kang and Rongsheng E. Wang*,&nbsp;","doi":"10.1021/acsbiomedchemau.4c0008410.1021/acsbiomedchemau.4c00084","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00084https://doi.org/10.1021/acsbiomedchemau.4c00084","url":null,"abstract":"<p >Dysregulated sialic acid biosynthesis is characteristic of the onset and progression of human diseases including hormone-sensitive prostate cancer and breast cancer. The sialylated glycoconjugates involved in this process are therefore important targets for identification and functional studies. To date, one of the most common strategies is metabolic glycoengineering, which utilizes <i>N</i>-acetylmannosamine (ManNAc) analogues such as <i>N</i>-azidoacetylmannosamine (ManNAz) to hijack sialic acid biosynthesis and label the sialylated glycoconjugates with “click chemistry (CuAAC)” tags. Yet, current chemical modifications including those CuAAC-based alkyne/azide tags are still big in size, and the resulting steric hindrance perturbs the mannosamine and sialic acid derivatives’ recognition and metabolism by enzymes involved in biosynthetic pathways. As a result, the peracetylated ManNAz has compromised incorporation to sialic acid substrates and manifests cellular growth inhibition and cytotoxicity. Herein, we show that the α-fluorinated peracetylated analogue ManN(F-Ac) displayed a satisfying safety profile in mammalian cell lines at concentrations as high as 500 μM. More importantly, aliphatic selenol-containing probes can efficiently displace α-fluorine in fluoroacetamide-containing substrates including ManN(F-Ac) at a neutral pH range (∼7.2). The combined use of peracetylated ManN(F-Ac) and the dethiobiotin-selenol probe as the fluorine-selenol displacement reaction (FSeDR) toolkit allowed for successful metabolic labeling of sialoglycoproteins in multiple prostate and cancer cell lines, including PC-3 and MDA-MB-231. More sialoglycoproteins in these cell lines were demonstrated to be labeled by FSeDR compared with the traditional CuAAC approach. Lastly, with FSeDR-mediated metabolic labeling, we were able to probe the cellular expression level and spatial distribution of sialylated glycoconjugates during the progression of these hormone-sensitive cancer cells. Taken together, the promising results suggest the potential of the FSeDR strategy to efficiently and systematically identify and study sialic acid substrates and potentially empower metabolic engineering on a diverse set of glycosylated proteins that are vital for human diseases.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"119–130 119–130"},"PeriodicalIF":3.8,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00084","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435956","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and Modification of Cordycepin-Phosphoramidate ProTide Derivatives for Antiviral Activity and Metabolic Stability","authors":"Aticha Thiraporn,&nbsp;Thanat Tiyasakulchai,&nbsp;Thitiphong Khamkhenshorngphanuch,&nbsp;Marie Hoarau,&nbsp;Ratthiya Thiabma,&nbsp;Suppachoke Onnome,&nbsp;Amporn Suphatrakul,&nbsp;Jaraspim Narkpuk,&nbsp;Chanya Srisaowakarn,&nbsp;Suwimon Manopwisedjaroen,&nbsp;Kitlada Srichomthong,&nbsp;Suradej Hongeng,&nbsp;Arunee Thitithanyanont,&nbsp;Peera Jaru-ampornpan,&nbsp;Sewan Theeramunkong,&nbsp;Bunpote Siridechadilok and Nitipol Srimongkolpithak*,&nbsp;","doi":"10.1021/acsbiomedchemau.4c0007110.1021/acsbiomedchemau.4c00071","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00071https://doi.org/10.1021/acsbiomedchemau.4c00071","url":null,"abstract":"<p >In our study, Prodrug nucleoTide (ProTide) technology was applied to cordycepin to enhance its antiviral activities and metabolic stability. Using cordycepin as starting material, we developed a synthetic method to access a series of stereospecific-phosphoramidate derivatives with various ester groups. We also successfully synthesized halogenated cordycepin derivatives via stannylation. Our 17 ProTide-cordycepin derivatives were pharmacologically evaluated for their antiviral activities. Phosphorus diastereomers <b>22</b><i>S</i> and <b>22</b><i>R</i> showed moderate inhibitory activity against corona and influenza viruses, while these compounds and derivatives (<b>25</b><i>S</i>, <b>27</b><i>S</i>, and <b>27</b><i>R</i>) demonstrated promising antiviral efficacy against dengue virus. Pharmacological screening indicated that <i>S</i>p-isomers generally exhibited slightly greater inhibitory activity than their <i>R</i>p-isomer counterparts against the dengue virus. The selected ProTides were assessed for their metabolic mechanism and stability via carboxypeptidase and microsomes. The hydrolysis rate of the <i>R</i>p-isomers was observed to be slightly higher than that of the <i>S</i>p-isomers, and the addition of a fluorine group also modestly increased this rate and fluorinated <b>39</b><i>S</i> extended its half-life compared to nonfluorinated counterparts. These findings suggested not only structure–activity relationships of cordycepin ProTide but also the comprehensive synthetic route to access cordycepin derivatives for further antiviral development.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"89–105 89–105"},"PeriodicalIF":3.8,"publicationDate":"2024-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00071","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and Modification of Cordycepin-Phosphoramidate ProTide Derivatives for Antiviral Activity and Metabolic Stability.","authors":"Aticha Thiraporn, Thanat Tiyasakulchai, Thitiphong Khamkhenshorngphanuch, Marie Hoarau, Ratthiya Thiabma, Suppachoke Onnome, Amporn Suphatrakul, Jaraspim Narkpuk, Chanya Srisaowakarn, Suwimon Manopwisedjaroen, Kitlada Srichomthong, Suradej Hongeng, Arunee Thitithanyanont, Peera Jaru-Ampornpan, Sewan Theeramunkong, Bunpote Siridechadilok, Nitipol Srimongkolpithak","doi":"10.1021/acsbiomedchemau.4c00071","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00071","url":null,"abstract":"<p><p>In our study, Prodrug nucleoTide (ProTide) technology was applied to cordycepin to enhance its antiviral activities and metabolic stability. Using cordycepin as starting material, we developed a synthetic method to access a series of stereospecific-phosphoramidate derivatives with various ester groups. We also successfully synthesized halogenated cordycepin derivatives via stannylation. Our 17 ProTide-cordycepin derivatives were pharmacologically evaluated for their antiviral activities. Phosphorus diastereomers <b>22</b> <i>S</i> and <b>22</b> <i>R</i> showed moderate inhibitory activity against corona and influenza viruses, while these compounds and derivatives (<b>25</b> <i>S</i>, <b>27</b> <i>S</i>, and <b>27</b> <i>R</i>) demonstrated promising antiviral efficacy against dengue virus. Pharmacological screening indicated that <i>S</i>p-isomers generally exhibited slightly greater inhibitory activity than their <i>R</i>p-isomer counterparts against the dengue virus. The selected ProTides were assessed for their metabolic mechanism and stability via carboxypeptidase and microsomes. The hydrolysis rate of the <i>R</i>p-isomers was observed to be slightly higher than that of the <i>S</i>p-isomers, and the addition of a fluorine group also modestly increased this rate and fluorinated <b>39</b> <i>S</i> extended its half-life compared to nonfluorinated counterparts. These findings suggested not only structure-activity relationships of cordycepin ProTide but also the comprehensive synthetic route to access cordycepin derivatives for further antiviral development.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"89-105"},"PeriodicalIF":3.8,"publicationDate":"2024-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843340/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metal-Triggered FAD Reduction in d-2-Hydroxyglutarate Dehydrogenase from <i>Pseudomonas aeruginosa</i> PAO1.","authors":"Joanna Afokai Quaye, Giovanni Gadda","doi":"10.1021/acsbiomedchemau.4c00108","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00108","url":null,"abstract":"<p><p>Alcohol oxidation is an indispensable chemical reaction in biological systems. This process, biologically catalyzed by alcohol dehydrogenases (ADHs) and alcohol oxidases (AOXs), follows two distinct chemical routes depending on the cofactor. ADHs have been widely demonstrated to require Zn²⁺- and NAD(P)⁺-based cosubstrates. Except for galactose oxidase, AOXs achieve their conversion of alcohols to aldehydes or ketones using flavin-based cofactors. The FMN-dependent α-hydroxy acid-oxidizing enzymes and the glucose-methanol-choline (GMC) superfamily abstract their substrate's α-OH proton using a catalytic histidine, leading to substrate oxidation and flavin reduction. However, there is no known alcohol oxidation mechanism for enzymes requiring both a flavin and a metal. The <i>Pseudomonas aeruginosa</i>d-2-hydroxyglutarate dehydrogenase (<i>Pa</i>D2HGDH) is a recently characterized α-hydroxy acid dehydrogenase that converts d-2-hydroxyglutarate or d-malate to 2-ketoglutarate or oxaloacetate, respectively. <i>Pa</i>D2HGDH requires FAD and Zn²⁺ for catalysis. Previous studies on <i>Pa</i>D2HGDH have identified a highly conserved active site histidine residue whose position is topologically conserved for catalytic bases in FMN-dependent α-hydroxy acid-oxidizing enzymes and the GMC superfamily of oxidoreductases. In this study, solvent isotope effects (SIEs) coupled with pL-rate profiles and a viscosity control have been used to probe the role of the Zn²⁺ cofactor in the C²-OH oxidation of d-malate and flavin reduction of <i>Pa</i>D2HGDH. The data revealed an inverse solvent equilibrium isotope effect (SEIE) of 0.51 ± 0.09 consistent with a Zn²⁺-triggered abstraction of the substrate C²-OH proton that initiates d-malate oxidation and flavin reduction. The system provides insights into the role of Zn²⁺ in the oxidation mechanism of <i>Pa</i>D2HGDH and, by extension, metallo flavoprotein dehydrogenases.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"204-214"},"PeriodicalIF":3.8,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843331/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metal-Triggered FAD Reduction in d-2-Hydroxyglutarate Dehydrogenase from Pseudomonas aeruginosa PAO1","authors":"Joanna Afokai Quaye,&nbsp; and ,&nbsp;Giovanni Gadda*,&nbsp;","doi":"10.1021/acsbiomedchemau.4c0010810.1021/acsbiomedchemau.4c00108","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00108https://doi.org/10.1021/acsbiomedchemau.4c00108","url":null,"abstract":"<p >Alcohol oxidation is an indispensable chemical reaction in biological systems. This process, biologically catalyzed by alcohol dehydrogenases (ADHs) and alcohol oxidases (AOXs), follows two distinct chemical routes depending on the cofactor. ADHs have been widely demonstrated to require Zn²⁺- and NAD(P)⁺-based cosubstrates. Except for galactose oxidase, AOXs achieve their conversion of alcohols to aldehydes or ketones using flavin-based cofactors. The FMN-dependent α-hydroxy acid-oxidizing enzymes and the glucose–methanol–choline (GMC) superfamily abstract their substrate’s α–OH proton using a catalytic histidine, leading to substrate oxidation and flavin reduction. However, there is no known alcohol oxidation mechanism for enzymes requiring both a flavin and a metal. The <i>Pseudomonas aeruginosa</i> <span>d</span>-2-hydroxyglutarate dehydrogenase (<i>Pa</i>D2HGDH) is a recently characterized α-hydroxy acid dehydrogenase that converts <span>d</span>-2-hydroxyglutarate or <span>d</span>-malate to 2-ketoglutarate or oxaloacetate, respectively. <i>Pa</i>D2HGDH requires FAD and Zn²⁺ for catalysis. Previous studies on <i>Pa</i>D2HGDH have identified a highly conserved active site histidine residue whose position is topologically conserved for catalytic bases in FMN-dependent α-hydroxy acid-oxidizing enzymes and the GMC superfamily of oxidoreductases. In this study, solvent isotope effects (SIEs) coupled with pL-rate profiles and a viscosity control have been used to probe the role of the Zn²⁺ cofactor in the C²–OH oxidation of <span>d</span>-malate and flavin reduction of <i>Pa</i>D2HGDH. The data revealed an inverse solvent equilibrium isotope effect (SEIE) of 0.51 ± 0.09 consistent with a Zn²⁺-triggered abstraction of the substrate C²–OH proton that initiates <span>d</span>-malate oxidation and flavin reduction. The system provides insights into the role of Zn²⁺ in the oxidation mechanism of <i>Pa</i>D2HGDH and, by extension, metallo flavoprotein dehydrogenases.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 1","pages":"204–214 204–214"},"PeriodicalIF":3.8,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00108","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435889","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}