Archives of biochemistry and biophysics最新文献_第8页

Degrading mutant IDH1 employing a PROTAC-based approach impairs STAT3 activation 使用基于protac的方法降解突变体IDH1损害STAT3激活。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-17 DOI: 10.1016/j.abb.2024.110281

Hashnu Dutta , Nishant Jain

Heterozygous mutations in IDH1 (isocitrate dehydrogenase 1) are found in most grade II and III brain tumors. A slew of mutant IDH1 inhibitors were identified soon after the discovery of IDH1 mutations in brain tumors. But recent reports show that mutant IDH1 inhibitors reverse therapeutic vulnerabilities and activate the oncogenic transcription factor STAT3 in mutant IDH1-expressing cells. Thus, inhibiting mutant IDH1 using mutant IDH1-specific inhibitors can result in drug resistance. Therefore, to block mutant IDH1, it is imperative to identify alternative modes of therapy. In these lines, recent findings show that PROteolysis TArgeting Chimera (PROTAC) molecules can be designed to degrade target proteins in cancer cells. However, it is unknown whether degrading mutant IDH1 leads to STAT3 activation. Therefore, in this study, we asked if degrading mutant IDH1 by employing a PROTAC-based approach leads to STAT3 activation. To answer the question, we adopted the dTAG system, where we fused FKBP12^F36V to mutant IDH1 proteins and used the FKBP12^F36V-specific PROTAC, dTAG-13, to degrade mutant IDH1-FKBP12^F36V. We assessed STAT3 activation in dTAG-13-treated cells expressing mutant IDH1-FKBP12^F36V. We found that fusing FKBP12^F36V-HA to mutant IDH1 phenocopies mutant IDH1 with similar expression levels, enzyme activity, and cellular localization. We observed that dTAG-13 degrades mutant IDH1-FKBP12^F36V-HA in a dose- and time-responsive manner. Unlike inhibiting, degrading mutant IDH1-FKBP12^F36V-HA did not lead to pSTAT3-Y705 activation. We conclude that degrading mutant IDH1 by employing a PROTAC-based approach impairs STAT3 activation. Based on these observations, we suggest that mutant IDH1-specific PROTACs can be developed to degrade mutant IDH1 in gliomas.

IDH1（异柠檬酸脱氢酶1）的杂合突变在大多数II级和III级脑肿瘤中发现。在脑肿瘤中发现IDH1突变后不久，就发现了大量突变的IDH1抑制剂。但最近的报道显示，突变型IDH1抑制剂逆转了治疗脆弱性，并激活了表达IDH1的突变细胞中的致癌转录因子STAT3。因此，使用IDH1突变体特异性抑制剂抑制IDH1突变体可导致耐药性。因此，为了阻断突变的IDH1，必须确定替代的治疗模式。在这些方面，最近的研究结果表明，靶向嵌合体的蛋白水解（PROTAC）分子可以被设计来降解癌细胞中的靶蛋白。然而，降解突变体IDH1是否会导致STAT3激活尚不清楚。因此，在本研究中，我们询问通过采用基于protac的方法降解突变体IDH1是否会导致STAT3激活。为了回答这个问题，我们采用了dTAG系统，将FKBP12F36V与突变体IDH1蛋白融合，并使用FKBP12F36V特异性PROTAC dTAG-13来降解突变体IDH1-FKBP12F36V。我们评估了表达突变体IDH1-FKBP12F36V的dtag -13处理细胞中STAT3的激活情况。我们发现，将FKBP12F36V-HA与突变体IDH1融合后，突变体IDH1具有相似的表达水平、酶活性和细胞定位。我们观察到dTAG-13以剂量和时间响应的方式降解突变体IDH1-FKBP12F36V-HA。与抑制不同，降解突变体IDH1-FKBP12F36V-HA不会导致pSTAT3-Y705激活。我们得出结论，采用基于protac的方法降解突变体IDH1会损害STAT3的激活。基于这些观察结果，我们建议可以开发突变型IDH1特异性PROTACs来降解胶质瘤中的突变型IDH1。

{"title":"Degrading mutant IDH1 employing a PROTAC-based approach impairs STAT3 activation","authors":"Hashnu Dutta , Nishant Jain","doi":"10.1016/j.abb.2024.110281","DOIUrl":"10.1016/j.abb.2024.110281","url":null,"abstract":"<div><div>Heterozygous mutations in IDH1 (isocitrate dehydrogenase 1) are found in most grade II and III brain tumors. A slew of mutant IDH1 inhibitors were identified soon after the discovery of IDH1 mutations in brain tumors. But recent reports show that mutant IDH1 inhibitors reverse therapeutic vulnerabilities and activate the oncogenic transcription factor STAT3 in mutant IDH1-expressing cells. Thus, inhibiting mutant IDH1 using mutant IDH1-specific inhibitors can result in drug resistance. Therefore, to block mutant IDH1, it is imperative to identify alternative modes of therapy. In these lines, recent findings show that <u>PRO</u>teolysis <u>TA</u>rgeting <u>C</u>himera (PROTAC) molecules can be designed to degrade target proteins in cancer cells. However, it is unknown whether degrading mutant IDH1 leads to STAT3 activation. Therefore, in this study, we asked if degrading mutant IDH1 by employing a PROTAC-based approach leads to STAT3 activation. To answer the question, we adopted the dTAG system, where we fused FKBP12<sup>F36V</sup> to mutant IDH1 proteins and used the FKBP12<sup>F36V</sup>-specific PROTAC, dTAG-13, to degrade mutant IDH1-FKBP12<sup>F36V</sup>. We assessed STAT3 activation in dTAG-13-treated cells expressing mutant IDH1-FKBP12<sup>F36V</sup>. We found that fusing FKBP12<sup>F36V</sup>-HA to mutant IDH1 phenocopies mutant IDH1 with similar expression levels, enzyme activity, and cellular localization. We observed that dTAG-13 degrades mutant IDH1-FKBP12<sup>F36V</sup>-HA in a dose- and time-responsive manner. Unlike inhibiting, degrading mutant IDH1-FKBP12<sup>F36V</sup>-HA did not lead to pSTAT3-Y705 activation. We conclude that degrading mutant IDH1 by employing a PROTAC-based approach impairs STAT3 activation. Based on these observations, we suggest that mutant IDH1-specific PROTACs can be developed to degrade mutant IDH1 in gliomas.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110281"},"PeriodicalIF":3.8,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mechanisms and applications of bacterial luciferase and its auxiliary enzymes 细菌荧光素酶及其辅助酶的作用机制及应用。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-15 DOI: 10.1016/j.abb.2025.110307

Chadaporn Kantiwiriyawanitch , Ubolsree Leartsakulpanich , Pimchai Chaiyen , Ruchanok Tinikul

Bacterial luciferase (LuxAB) catalyzes the conversion of reduced flavin mononucleotide (FMNH⁻), oxygen, and a long-chain aldehyde to oxidized FMN, the corresponding acid and water with concomitant light emission. This bioluminescence reaction requires the reaction of a flavin reductase such as LuxG (in vivo partner of LuxAB) to supply FMNH⁻ for the LuxAB reaction. LuxAB is a well-known self-sufficient luciferase system because both aldehyde and FMNH⁻ substrates can be produced by the associated enzymes encoded by the genes in the lux operon, allowing the system to be auto-luminous. This makes it useful for in vivo applications. Structural and functional studies have long been performed in efforts to gain a better understanding of the LuxAB reaction. Recently, continued exploration of the LuxAB reaction have elucidated the mechanisms of C4a-hydroperoxyflavin formation and identified key catalytic residues such as His44 that facilitates the generation of flavin intermediates important for light generation. Advancements in protein engineering and synthetic biology have improved the bioluminescence properties of LuxAB. Various applications of LuxAB for bioimaging, bioreporters, biosensing in metabolic engineering and real-time monitoring of aldehyde metabolites in biofuel production pathways have been developed during the last decade. Challenging issues such as achieving red-shifted emissions, optimizing the signal intensity and identifying mechanisms related to the generation of light-emitting species remain to be explored. Nevertheless, LuxAB continues to be a promising tool for diverse biotechnological and biomedical applications.

细菌荧光素酶（LuxAB）催化还原黄素单核苷酸（FMNH），氧和长链醛转化为氧化的FMN，相应的酸和水，并伴随发光。这种生物发光反应需要黄素还原酶如LuxG （LuxAB的体内伙伴）的反应来提供FMNH⁻（为LuxAB反应提供FMNH⁻）。LuxAB是一个众所周知的自给自足的荧光素酶系统，因为醛和FMNH -底物都可以由lux操纵子中基因编码的相关酶产生，从而使该系统能够自动发光。这使得它对体内应用很有用。长期以来，人们一直在进行结构和功能研究，以更好地了解LuxAB反应。最近，LuxAB反应的持续探索已经阐明了c4a -氢过氧黄素形成的机制，并确定了His44等促进黄素中间体生成的关键催化残基，这些中间体对光的产生很重要。蛋白质工程和合成生物学的进步改善了LuxAB的生物发光特性。在过去十年中，LuxAB在生物成像、生物报告、代谢工程中的生物传感和生物燃料生产途径中醛代谢物的实时监测等方面的各种应用得到了发展。诸如实现红移发射、优化信号强度以及确定与发光物种产生相关的机制等具有挑战性的问题仍有待探索。尽管如此，LuxAB仍然是各种生物技术和生物医学应用的有前途的工具。

{"title":"Mechanisms and applications of bacterial luciferase and its auxiliary enzymes","authors":"Chadaporn Kantiwiriyawanitch , Ubolsree Leartsakulpanich , Pimchai Chaiyen , Ruchanok Tinikul","doi":"10.1016/j.abb.2025.110307","DOIUrl":"10.1016/j.abb.2025.110307","url":null,"abstract":"<div><div>Bacterial luciferase (LuxAB) catalyzes the conversion of reduced flavin mononucleotide (FMNH⁻), oxygen, and a long-chain aldehyde to oxidized FMN, the corresponding acid and water with concomitant light emission. This bioluminescence reaction requires the reaction of a flavin reductase such as LuxG (<em>in vivo</em> partner of LuxAB) to supply FMNH⁻ for the LuxAB reaction. LuxAB is a well-known self-sufficient luciferase system because both aldehyde and FMNH⁻ substrates can be produced by the associated enzymes encoded by the genes in the <em>lux</em> operon, allowing the system to be auto-luminous. This makes it useful for <em>in vivo</em> applications. Structural and functional studies have long been performed in efforts to gain a better understanding of the LuxAB reaction. Recently, continued exploration of the LuxAB reaction have elucidated the mechanisms of C4a-hydroperoxyflavin formation and identified key catalytic residues such as His44 that facilitates the generation of flavin intermediates important for light generation. Advancements in protein engineering and synthetic biology have improved the bioluminescence properties of LuxAB. Various applications of LuxAB for bioimaging, bioreporters, biosensing in metabolic engineering and real-time monitoring of aldehyde metabolites in biofuel production pathways have been developed during the last decade. Challenging issues such as achieving red-shifted emissions, optimizing the signal intensity and identifying mechanisms related to the generation of light-emitting species remain to be explored. Nevertheless, LuxAB continues to be a promising tool for diverse biotechnological and biomedical applications.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110307"},"PeriodicalIF":3.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

c-FLIP/Ku70 complex; A potential molecular target for apoptosis induction in hepatocellular carcinoma c-FLIP / Ku70复杂;肝细胞癌诱导细胞凋亡的潜在分子靶点。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-14 DOI: 10.1016/j.abb.2025.110306

Yasamin Haghir-Sharif-Zamini , Arezoo Khosravi , Moustapha Hassan , Ali Zarrabi , Massoud Vosough

Hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide and the most common form of liver cancer. Despite global efforts toward early diagnosis and effective treatments, HCC is often diagnosed at advanced stages, where conventional therapies frequently lead to resistance and/or high recurrence rates. Therefore, novel biomarkers and promising medications are urgently required. Epi-drugs, or epigenetic-based medicines, have recently emerged as a promising therapeutic modality. Since the epigenome of the cancer cells is always dysregulated and this is followed by apoptosis-resistance, reprogramming the epigenome of cancer cells by epi-drugs (such as HDAC inhibitors (HDACis), and DNMT inhibitors (DNMTis)) could be an alternative approach to use in concert with established treatment protocols. C-FLIP, an anti-apoptotic protein, and Ku70, a member of the DNA repair system, bind together and make a cytoplasmic complex in certain cancers and induce resistance to apoptosis. Many epi-drugs, such as HDACis, can dissociate this complex through Ku70 acetylation and activate cellular apoptosis. The novel compounds for dissociating this complex could provide an innovative insight into molecular targeted HCC treatments. In this review, we address the innovative therapeutic potential of targeting c-FLIP/Ku70 complex by epi-drugs, particularly HDACis, to overcome apoptosis resistance of HCC cells. This review will cover the mechanisms by which the c-FLIP/Ku70 complex facilitates cancer cell survival, the impact of epigenetic alterations on the complex dissociation, and highlight HDACis potential in combination therapies, biomarker developments and mechanistic overviews. This review highlights c-FLIP ubiquitination and Ku70 acetylation levels as diagnostic and prognostic tools in HCC management.

肝细胞癌（HCC）是全球最致命的恶性肿瘤之一，也是最常见的肝癌形式。尽管全球致力于早期诊断和有效治疗，但HCC通常在晚期才被诊断出来，而常规治疗往往导致耐药和/或高复发率。因此，迫切需要新的生物标志物和有前景的药物。外显药物，或基于表观遗传学的药物，最近成为一种有前途的治疗方式。由于癌细胞的表观基因组总是失调，随之而来的是凋亡抵抗，因此通过表观药物（如HDAC抑制剂（HDACis）和DNMT抑制剂（DNMTis））对癌细胞的表观基因组进行重编程可能是与既定治疗方案相一致的另一种方法。抗凋亡蛋白C-FLIP和DNA修复系统成员Ku70结合在一起，在某些癌症中形成细胞质复合体，诱导细胞凋亡抵抗。许多外显药物，如HDACis，可以通过Ku70乙酰化解离该复合物并激活细胞凋亡。解离该复合物的新化合物可能为分子靶向HCC治疗提供创新的见解。在这篇综述中，我们讨论了用外显药物，特别是HDACis靶向c-FLIP/Ku70复合物，克服HCC细胞凋亡抵抗的创新治疗潜力。本文将涵盖c-FLIP/Ku70复合体促进癌细胞存活的机制，表观遗传改变对复合体解离的影响，并强调HDACis在联合治疗、生物标志物开发和机制概述方面的潜力。本综述强调c-FLIP泛素化和Ku70乙酰化水平是HCC治疗的诊断和预后工具。

{"title":"c-FLIP/Ku70 complex; A potential molecular target for apoptosis induction in hepatocellular carcinoma","authors":"Yasamin Haghir-Sharif-Zamini , Arezoo Khosravi , Moustapha Hassan , Ali Zarrabi , Massoud Vosough","doi":"10.1016/j.abb.2025.110306","DOIUrl":"10.1016/j.abb.2025.110306","url":null,"abstract":"<div><div>Hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide and the most common form of liver cancer. Despite global efforts toward early diagnosis and effective treatments, HCC is often diagnosed at advanced stages, where conventional therapies frequently lead to resistance and/or high recurrence rates. Therefore, novel biomarkers and promising medications are urgently required. Epi-drugs, or epigenetic-based medicines, have recently emerged as a promising therapeutic modality. Since the epigenome of the cancer cells is always dysregulated and this is followed by apoptosis-resistance, reprogramming the epigenome of cancer cells by epi-drugs (such as HDAC inhibitors (HDACis), and DNMT inhibitors (DNMTis)) could be an alternative approach to use in concert with established treatment protocols. C-FLIP, an anti-apoptotic protein, and Ku70, a member of the DNA repair system, bind together and make a cytoplasmic complex in certain cancers and induce resistance to apoptosis. Many epi-drugs, such as HDACis, can dissociate this complex through Ku70 acetylation and activate cellular apoptosis. The novel compounds for dissociating this complex could provide an innovative insight into molecular targeted HCC treatments. In this review, we address the innovative therapeutic potential of targeting c-FLIP/Ku70 complex by epi-drugs, particularly HDACis, to overcome apoptosis resistance of HCC cells. This review will cover the mechanisms by which the c-FLIP/Ku70 complex facilitates cancer cell survival, the impact of epigenetic alterations on the complex dissociation, and highlight HDACis potential in combination therapies, biomarker developments and mechanistic overviews. This review highlights c-FLIP ubiquitination and Ku70 acetylation levels as diagnostic and prognostic tools in HCC management.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110306"},"PeriodicalIF":3.8,"publicationDate":"2025-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Live-cell FRET assay on the stoichiometry and affinity of the YAP complexes in MCF-7 cells MCF-7细胞中YAP复合物的化学计量学和亲和力的活细胞FRET测定。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-14 DOI: 10.1016/j.abb.2025.110305

Yongtong Zhan , Lingao Dai , Ze Fu , Xuhong Fan , Xin Li , Guihao Wu , Yue Ni , Ge Wu , Tongsheng Chen , Xiaoping Wang

Yes-associated protein (YAP), a focal point of current biological research, is involved in regulating various life processes. In this report, live-cell fluorescence resonance energy transfer (FRET) imaging was employed to unravel the YAP complexes in MCF-7 cells. Fluorescence imaging of living cells co-expressing CFP (cyan fluorescent protein)-YAP and YFP (yellow fluorescent protein)-LATS1 (large tumor suppressor 1) plasmids revealed that YAP promoted LATS1 oligomerization around mitochondria. Moreover, FRET two-hybrid assay showed that YAP directly interacted with LATS1 to form dimer. Similarly, we found that YAP directly interacted with large tumor suppressor 2 (LATS2) to form a heterotrimer with 1:2 in cytoplasm and around mitochondria. In addition, YAP directly interacted with angiomotin (AMOT) to form a heterodimer in cytoplasm. However, YAP did not interact with O-linked N-acetylglucosamine transferase (OGT). Furthermore, FRET assay also indicated that YAP exhibited a higher affinity with AMOT, followed by LATS1, and least with LATS2. In summary, YAP directly interacts with LATS1 and AMOT to form a heterodimer, with LATS2 to form a heterotrimer with 1:2, and shows a preference for binding to AMOT, followed by LATS1, and lastly LATS2, providing new insights into the Hippo-YAP signaling pathway.

Yes-associated protein （YAP）是当前生物学研究的热点之一，它参与调节多种生命过程。在本报告中，利用活细胞荧光共振能量转移（FRET）成像来解开MCF-7细胞中的YAP复合物。共表达CFP（青色荧光蛋白）-YAP和YFP（黄色荧光蛋白）-LATS1（大肿瘤抑制因子1）质粒的活细胞荧光成像显示，YAP促进了线粒体周围LATS1的寡聚化。此外，FRET双杂交实验表明，YAP直接与LATS1相互作用形成二聚体。同样，我们发现YAP直接与大肿瘤抑制因子2 （LATS2）相互作用，在细胞质和线粒体周围形成1:2的异源三聚体。此外，YAP直接与血管运动素（AMOT）相互作用，在细胞质中形成异二聚体。然而，YAP不与O-linked N-acetylglucosamine transferase （OGT）相互作用。此外，FRET实验还表明，YAP与AMOT的亲和力较高，其次是LATS1，与LATS2的亲和力最低。综上所述，YAP直接与LATS1和AMOT相互作用形成异源二聚体，与LATS2形成1:2的异源三聚体，并且优先结合AMOT，其次是LATS1，最后是LATS2，这为Hippo-YAP信号通路提供了新的认识。

{"title":"Live-cell FRET assay on the stoichiometry and affinity of the YAP complexes in MCF-7 cells","authors":"Yongtong Zhan , Lingao Dai , Ze Fu , Xuhong Fan , Xin Li , Guihao Wu , Yue Ni , Ge Wu , Tongsheng Chen , Xiaoping Wang","doi":"10.1016/j.abb.2025.110305","DOIUrl":"10.1016/j.abb.2025.110305","url":null,"abstract":"<div><div>Yes-associated protein (YAP), a focal point of current biological research, is involved in regulating various life processes. In this report, live-cell fluorescence resonance energy transfer (FRET) imaging was employed to unravel the YAP complexes in MCF-7 cells. Fluorescence imaging of living cells co-expressing CFP (cyan fluorescent protein)-YAP and YFP (yellow fluorescent protein)-LATS1 (large tumor suppressor 1) plasmids revealed that YAP promoted LATS1 oligomerization around mitochondria. Moreover, FRET two-hybrid assay showed that YAP directly interacted with LATS1 to form dimer. Similarly, we found that YAP directly interacted with large tumor suppressor 2 (LATS2) to form a heterotrimer with 1:2 in cytoplasm and around mitochondria. In addition, YAP directly interacted with angiomotin (AMOT) to form a heterodimer in cytoplasm. However, YAP did not interact with O-linked N-acetylglucosamine transferase (OGT). Furthermore, FRET assay also indicated that YAP exhibited a higher affinity with AMOT, followed by LATS1, and least with LATS2. In summary, YAP directly interacts with LATS1 and AMOT to form a heterodimer, with LATS2 to form a heterotrimer with 1:2, and shows a preference for binding to AMOT, followed by LATS1, and lastly LATS2, providing new insights into the Hippo-YAP signaling pathway.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110305"},"PeriodicalIF":3.8,"publicationDate":"2025-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unveiling the cyclopropyl appended acyl thiourea derivatives as antimicrobial, α-amylase and proteinase K inhibitors: Design, synthesis, biological evaluation, molecular docking, DFT and ADMET studies 揭示环丙基附加酰基硫脲衍生物作为抗菌、α-淀粉酶和蛋白酶K抑制剂：设计、合成、生物学评价、分子对接、DFT和ADMET研究。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-13 DOI: 10.1016/j.abb.2025.110304

Hina Zaman , Aamer Saeed , Hammad Ismail , Muhammad Rashid

Acyl thiourea scaffolds are frequently employed in drug development to discern unique and essential therapies for the eradication of the most challenging diseases. Hence, we developed a library of novel cyclopropyl incorporating acyl thiourea derivatives (4a-j) and evaluated their antimicrobial, α-amylase, and proteinase K inhibition potential. Compound (4h) (4-methoxy) demonstrated the strongest α-amylase inhibition (IC₅₀ = 1.572 ± 0.017 μM), while compound (4j) (3,4,5-trimethoxy) exhibited potent proteinase K inhibition (IC₅₀ = 1.718 ± 0.061 μM), comparable to the standard acarbose (IC₅₀ = 1.063 ± 0.013 μM) and phenyl methyl sulfonyl fluoride (IC₅₀ = 0.119 ± 0.014 μM). The unsubstituted compound (4a) emerged as the most potent antifungal agent (17 mm zone of inhibition), outperforming the positive control Terbinafine (zone of inhibition 16 mm). These compounds (4a-j) also displayed moderate antibacterial activity. SAR analysis revealed the influences of various substitutions on the acyl thiourea scaffold. Computational studies, including DFT, molecular docking, and ADMET predictions, supported the biological findings and identified these compounds as promising inhibitors of α-amylase, proteinase K, and microbial pathogens.

酰基硫脲支架经常用于药物开发，以识别根除最具挑战性疾病的独特和必要的疗法。因此，我们建立了一个新的含酰基硫脲衍生物环丙基文库（4a-j），并评估了它们的抗菌、α-淀粉酶和蛋白酶K抑制潜力。化合物（4h）（4-甲氧基）对α-淀粉酶的抑制作用最强（IC50 = 1.572±0.017），而化合物（4j）（3,4,5-三甲氧基）对蛋白酶K的抑制作用最强（1.718±0.061 μM），与标准阿卡波糖（IC50 = 1.063±0.013 μM）和苯基甲基磺酰氟（IC50 = 0.119±0.014 μM）相当。未取代化合物（4a）成为最有效的抗真菌剂（17 mm抑制区），优于阳性对照特比萘芬（16 mm抑制区）。这些化合物（4a-j）也显示出中等的抗菌活性。SAR分析揭示了各种取代对酰基硫脲支架的影响。计算研究，包括DFT、分子对接和ADMET预测，支持生物学发现，并确定这些化合物是α-淀粉酶、蛋白酶K和微生物病原体的有希望的抑制剂。

{"title":"Unveiling the cyclopropyl appended acyl thiourea derivatives as antimicrobial, α-amylase and proteinase K inhibitors: Design, synthesis, biological evaluation, molecular docking, DFT and ADMET studies","authors":"Hina Zaman , Aamer Saeed , Hammad Ismail , Muhammad Rashid","doi":"10.1016/j.abb.2025.110304","DOIUrl":"10.1016/j.abb.2025.110304","url":null,"abstract":"<div><div>Acyl thiourea scaffolds are frequently employed in drug development to discern unique and essential therapies for the eradication of the most challenging diseases. Hence, we developed a library of novel cyclopropyl incorporating acyl thiourea derivatives (<strong>4a-j</strong>) and evaluated their antimicrobial, <em>α</em>-amylase, and proteinase K inhibition potential. Compound (<strong>4h</strong>) (4-methoxy) demonstrated the strongest <em>α</em>-amylase inhibition (IC<sub>50</sub> = 1.572 ± 0.017 μM), while compound (<strong>4j</strong>) (3,4,5-trimethoxy) exhibited potent proteinase K inhibition (IC<sub>50</sub> = 1.718 ± 0.061 μM), comparable to the standard acarbose (IC<sub>50</sub> = 1.063 ± 0.013 μM) and phenyl methyl sulfonyl fluoride (IC<sub>50</sub> = 0.119 ± 0.014 μM). The unsubstituted compound (<strong>4a</strong>) emerged as the most potent antifungal agent (17 mm zone of inhibition), outperforming the positive control Terbinafine (zone of inhibition 16 mm). These compounds (<strong>4a-j</strong>) also displayed moderate antibacterial activity. SAR analysis revealed the influences of various substitutions on the acyl thiourea scaffold. Computational studies, including DFT, molecular docking, and ADMET predictions, supported the biological findings and identified these compounds as promising inhibitors of <em>α</em>-amylase, proteinase K, and microbial pathogens.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110304"},"PeriodicalIF":3.8,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Gymnema saponin-induced lipid flip-flop identifies rigid membrane phenotype of methicillin resistant S. aureus and enhances it's antibiotic susceptibility 匙羹藤皂苷诱导的脂质翻转鉴定耐甲氧西林金黄色葡萄球菌刚性膜表型并增强其抗生素敏感性。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-11 DOI: 10.1016/j.abb.2025.110303

Gayatree Panda , Swagatika Dehury , Himadri Gourav Behuria , Bijesh Kumar Biswal , Ashis Kumar Jena , Indrani Mohanty , Sasmita Hotta , Santosh Kumar Padhi , Santosh Kumar Sahu

Our previous study revealed that lipid flip-flop inducing phytochemicals from Gymnema sylvestre increase membrane permeability of antimicrobials in S. aureus. However, their lipid flipping and membrane permeabilizing effect on methicillin resistant S. aureus (MRSA) membrane that has intrinsically higher aminoacylated lipid content compared to methicillin sensitive S. aureus (MSSA) is poorly characterized. Gymnema saponins, gymnemic acid I and IV significantly increased the antibiotic susceptibility in both MSSA and MRSA. MRSA exhibited a rigid membrane with lipid diffusion coefficient 0.0002 μm²/s compared to the MSSA membrane lipids with diffusion coefficient 1.48 μm²/s. Further, unlike MSSA, MRSA cells inhibited fusion of fluid liposomes with their plasma membrane. In vitro assay on reconstituted membrane vesicles revealed that Gymnema saponins induced 60 % lipid flipping in MSSA membrane compared to only 20 % lipid flipping in MRSA, indicating significantly lower Gymnema saponin-induced trans-bilayer lipid mobility in MRSA. Gymnema saponins induced significantly lower crystal violet uptake, release of cellular protein, cell shrinkage and lysis in MRSA compared to MSSA. Gymnema saponins led to dose-dependent inhibition of lipid-aminoacylation in both MSSA and MRSA making their membranes more negative compared to untreated control cells. In silico analysis reveals binding of both gymnemic acid I and IV to multiple peptide resistance factor (binding energy ∼ 7.5 kCal), the protein responsible for lipid aminoacylation in S. aureus. For the first time, our study reveals that MRSA membrane with higher aminoacyl-PG compared to MSSA shows significantly lower rate of diffusion and trans-bilayer flip-flop of lipids. Further, gymnemic acids are useful probes for identification, characterization and drug sensitization of rigid membrane MRSA phenotypes.

我们之前的研究表明，脂质翻转诱导的植物化学物质来自金螺旋体，增加了金黄色葡萄球菌抗微生物药物的膜通透性。然而，与甲氧西林敏感金黄色葡萄球菌（MSSA）相比，它们对耐甲氧西林金黄色葡萄球菌（MRSA）具有更高氨酰化脂质含量的膜的脂质翻转和膜透性作用尚不清楚。裸子皂苷、裸子酸I和IV显著增加了msa和MRSA的抗生素敏感性。MRSA膜脂质扩散系数为0.0002 μm2/s，而MSSA膜脂质扩散系数为1.48 μm2/s。此外，与MSSA不同，MRSA细胞抑制流体脂质体与其质膜的融合。重组膜囊的体外实验显示，木藤皂苷在MSSA膜中诱导60%的脂质翻转，而在MRSA膜中仅诱导20%的脂质翻转，这表明木藤皂苷诱导的MRSA跨双层脂质流动性明显降低。与MSSA相比，藤皂苷诱导MRSA的结晶紫吸收、细胞蛋白释放、细胞收缩和裂解显著降低。裸子皂苷导致MSSA和MRSA中脂质氨基酰化的剂量依赖性抑制，使其膜比未处理的对照细胞更阴性。硅分析显示，裸子酸I和IV与多肽抗性因子（结合能~ 7.5 kCal）结合，该蛋白在金黄色葡萄球菌中负责脂质氨基酰化。我们的研究首次揭示了与MSSA相比，具有较高氨基酰基pg的MRSA膜具有明显较低的扩散速率和脂质跨双层翻转。此外，裸子酸是鉴定、表征和药物致敏刚性膜MRSA表型的有用探针。

{"title":"Gymnema saponin-induced lipid flip-flop identifies rigid membrane phenotype of methicillin resistant S. aureus and enhances it's antibiotic susceptibility","authors":"Gayatree Panda , Swagatika Dehury , Himadri Gourav Behuria , Bijesh Kumar Biswal , Ashis Kumar Jena , Indrani Mohanty , Sasmita Hotta , Santosh Kumar Padhi , Santosh Kumar Sahu","doi":"10.1016/j.abb.2025.110303","DOIUrl":"10.1016/j.abb.2025.110303","url":null,"abstract":"<div><div>Our previous study revealed that lipid flip-flop inducing phytochemicals from <em>Gymnema sylvestre</em> increase membrane permeability of antimicrobials in <em>S</em>. <em>aureus</em>. However, their lipid flipping and membrane permeabilizing effect on methicillin resistant <em>S</em>. <em>aureus</em> (MRSA) membrane that has intrinsically higher aminoacylated lipid content compared to methicillin sensitive <em>S</em>. <em>aureus</em> (MSSA) is poorly characterized. <em>Gymnema</em> saponins, gymnemic acid I and IV significantly increased the antibiotic susceptibility in both MSSA and MRSA. MRSA exhibited a rigid membrane with lipid diffusion coefficient 0.0002 μm<sup>2</sup>/s compared to the MSSA membrane lipids with diffusion coefficient 1.48 μm<sup>2</sup>/s. Further, unlike MSSA, MRSA cells inhibited fusion of fluid liposomes with their plasma membrane. <em>In vitro</em> assay on reconstituted membrane vesicles revealed that <em>Gymnema</em> saponins induced 60 % lipid flipping in MSSA membrane compared to only 20 % lipid flipping in MRSA, indicating significantly lower <em>Gymnema</em> saponin-induced <em>trans</em>-bilayer lipid mobility in MRSA. <em>Gymnema</em> saponins induced significantly lower crystal violet uptake, release of cellular protein, cell shrinkage and lysis in MRSA compared to MSSA. <em>Gymnema</em> saponins led to dose-dependent inhibition of lipid-aminoacylation in both MSSA and MRSA making their membranes more negative compared to untreated control cells. <em>In silico</em> analysis reveals binding of both gymnemic acid I and IV to multiple peptide resistance factor (binding energy ∼ 7.5 kCal), the protein responsible for lipid aminoacylation in <em>S</em>. <em>aureus</em>. For the first time, our study reveals that MRSA membrane with higher aminoacyl-PG compared to MSSA shows significantly lower rate of diffusion and <em>trans</em>-bilayer flip-flop of lipids. Further, gymnemic acids are useful probes for identification, characterization and drug sensitization of rigid membrane MRSA phenotypes.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110303"},"PeriodicalIF":3.8,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142977294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The effectiveness of prolonged hypothermic preservation of isolated rat hearts using oxygen, medical nitrous oxide and carbon monoxide gas mixtures 用氧气、医用氧化亚氮和一氧化碳混合气体对离体大鼠心脏进行长期低温保存的效果。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-10 DOI: 10.1016/j.abb.2025.110295

Evgeniy L. Gagarinskiy, Mars G. Sharapov, Ruslan G. Goncharov, Artem E. Gurin, Svetlana V. Ugraitskaya, Eugeny E. Fesenko

The possibility of using an oxygen-nitrous oxide mixture for prolonged hypothermic preservation of rat heart for 24 h was investigated. A comparative analysis of restoration of functional activity of hearts in the groups of 24-h preservation at +4 °C with different gases (O₂, N₂) and gas mixtures (CO + O₂, N₂O + O₂, N₂+O₂, N₂O + N₂) was carried out. It was shown that the presence of oxygen in the gas mixture was the key factor for heart preservation. No stable heart preservation was observed in oxygen-free mixtures. At the same time, preservation in pure oxygen showed a significantly lower level of cardiac recovery compared to preservation in gas mixtures O₂+CO (6.5 atm.) and O₂+N₂O (6.5 atm.). LVDP (left ventricular developed pressure) values were 30 ± 19 mmHg and 46 ± 9 mmHg, respectively, with no significant differences found. The decrease in LDVP after 24 h of storage was 26–40 % of the intact control. The results obtained indicate the presence of pronounced synergistic effects of both gases during 24-h heart preservation, which is confirmed by data of marker genes Nfe2l2, Nox1, Prdx1, Hif1a, Nos2, Slc2a4, Ucp-1, Jun, Casp3 expression analysis and myocardial infarction damage level data. The more frequent occurrence of arrhythmias was observed in the oxygen-nitrous oxide group compared with the CO group, and the mechanism of this phenomenon is unclear. Nevertheless, the already medically approved N₂O + O₂ gas mixture could serve as a balanced choice for future improvements, offering a shorter duration of cardiac preservation compared to the CO + O₂ mixture, while ensuring safety in its use.

探讨了氧-氧化亚氮混合物延长大鼠心脏低温保存24小时的可能性。比较分析不同气体（O2、N2）和混合气体（CO+O2、N2O+O2、N2+O2、N2O+N2） +4℃24小时保存组心脏功能活动恢复情况。结果表明，混合气体中氧气的存在是心脏保存的关键因素。在无氧混合物中没有观察到稳定的心脏保存。同时，与保存在O2+CO （6.5 atm.）和O2+N2O （6.5 atm.）混合气体中相比，保存在纯氧中的心脏恢复水平明显较低。左心室发育压（LVDP）值分别为30±19 mmHg和46±9 mmHg，两组间无显著差异。贮藏24小时后LDVP下降幅度为完整对照的26-40%。结果表明，两种气体在24小时心脏保存过程中存在明显的协同作用，这一结果得到了标记基因Nfe2l2、Nox1、Prdx1、Hif1a、Nos2、Slc2a4、Ucp-1、Jun、Casp3表达分析和心肌梗死损伤水平数据的证实。氧-氧化亚氮组心律失常发生率高于一氧化碳组，其机制尚不清楚。尽管如此，医学上已经批准的N2O+O2气体混合物可以作为未来改进的平衡选择，与CO+O2混合物相比，提供更短的心脏保存时间，同时确保其使用的安全性。

{"title":"The effectiveness of prolonged hypothermic preservation of isolated rat hearts using oxygen, medical nitrous oxide and carbon monoxide gas mixtures","authors":"Evgeniy L. Gagarinskiy, Mars G. Sharapov, Ruslan G. Goncharov, Artem E. Gurin, Svetlana V. Ugraitskaya, Eugeny E. Fesenko","doi":"10.1016/j.abb.2025.110295","DOIUrl":"10.1016/j.abb.2025.110295","url":null,"abstract":"<div><div>The possibility of using an oxygen-nitrous oxide mixture for prolonged hypothermic preservation of rat heart for 24 h was investigated. A comparative analysis of restoration of functional activity of hearts in the groups of 24-h preservation at +4 °C with different gases (O<sub>2</sub>, N<sub>2</sub>) and gas mixtures (CO + O<sub>2</sub>, N<sub>2</sub>O + O<sub>2</sub>, N<sub>2</sub>+O<sub>2</sub>, N<sub>2</sub>O + N<sub>2</sub>) was carried out. It was shown that the presence of oxygen in the gas mixture was the key factor for heart preservation. No stable heart preservation was observed in oxygen-free mixtures. At the same time, preservation in pure oxygen showed a significantly lower level of cardiac recovery compared to preservation in gas mixtures O<sub>2</sub>+CO (6.5 atm.) and O<sub>2</sub>+N<sub>2</sub>O (6.5 atm.). LVDP (left ventricular developed pressure) values were 30 ± 19 mmHg and 46 ± 9 mmHg, respectively, with no significant differences found. The decrease in LDVP after 24 h of storage was 26–40 % of the intact control. The results obtained indicate the presence of pronounced synergistic effects of both gases during 24-h heart preservation, which is confirmed by data of marker genes Nfe2l2, Nox1, Prdx1, Hif1a, Nos2, Slc2a4, Ucp-1, Jun, Casp3 expression analysis and myocardial infarction damage level data. The more frequent occurrence of arrhythmias was observed in the oxygen-nitrous oxide group compared with the CO group, and the mechanism of this phenomenon is unclear. Nevertheless, the already medically approved N<sub>2</sub>O + O<sub>2</sub> gas mixture could serve as a balanced choice for future improvements, offering a shorter duration of cardiac preservation compared to the CO + O<sub>2</sub> mixture, while ensuring safety in its use.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"765 ","pages":"Article 110295"},"PeriodicalIF":3.8,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142969503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

DANCR knockdown alleviates neuroinflammation and functional recovery after spinal cord injury via regulating the ACTN4 / STAT3 axis danr敲低通过调节ACTN4 / STAT3轴减轻脊髓损伤后神经炎症和功能恢复。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-09 DOI: 10.1016/j.abb.2025.110293

Bin Xia , Cheng Yu , Jin Liu , Jiezhao Lin , Jiao Lyu , Xin Wang , Lixin Zhu

Polarization of microglia following spinal cord injury (SCI) is a pivotal pathological process of secondary injury. Although differentiation antagonistic nonprotein coding RNA (DANCR) has been implicated in immune and inflammatory responses across various diseases, its role in SCI still unclear. This research aimed to clarify the underlying mechanisms of DANCR in SCI recovery by investigating its expression pattern in microglia. Our findings indicate that the DANCR level in microglia is increased after SCI and that its knockdown can promote microglial M2-type polarization; suppress inflammatory cytokines, oxidative stress, and neuronal apoptosis; and facilitate nerve regeneration as well as spinal cord functional recovery. Further investigations suggest that DANCR's effects are mediated through the ACTN4/STAT3 axis. These results provide potential targets for enhancing functional recovery following SCI.

脊髓损伤后小胶质细胞的极化是继发性损伤的关键病理过程。尽管分化拮抗非蛋白编码RNA （DANCR）与多种疾病的免疫和炎症反应有关，但其在脊髓损伤中的作用尚不清楚。本研究旨在通过研究DANCR在小胶质细胞中的表达模式来阐明其在脊髓损伤恢复中的潜在机制。我们的研究结果表明，脊髓损伤后小胶质细胞中的DANCR水平升高，其下调可促进小胶质细胞m2型极化；抑制炎症细胞因子、氧化应激和神经元凋亡；促进神经再生和脊髓功能恢复。进一步的研究表明，DANCR的作用是通过ACTN4/STAT3轴介导的。这些结果为增强脊髓损伤后的功能恢复提供了潜在的靶点。

引用次数: 0

Assessing the role of Berberine as an inhibitor of advanced glycation end products (AGEs) formation using in vitro and molecular interaction studies 利用体外和分子相互作用研究评估小檗碱作为晚期糖基化终产物（AGEs）形成抑制剂的作用。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-09 DOI: 10.1016/j.abb.2025.110292

Yusra Ahmad , Faisal Nabi , Sana Siddiqui , Rizwan Hasan Khan , Safia Habib , Shagufta Moin

Glycation leads to the formation of protein aggregates and advanced glycation end products (AGEs) by non-enzymatic reaction. AGEs have been linked to several pathological conditions such as diabetes, cardiovascular disorders, Alzheimer's etc. Our research objective is understanding how methylglyoxal triggers AGEs and protein aggregate formation in human serum albumin (HSA) and how the phytochemical berberine protects it. Employing various biochemical and biophysical techniques, we explored how berberine alters human serum albumin's biochemical properties and structure during multiple glycation stages. HSA was incubated with methylglyoxal at varying concentrations of berberine for 7–14 days at a temperature range of 35–37 degrees C. Methylglyoxal induced the formation of AGEs, fibrillar aggregates and hydrophobic protein patches in HSA as demonstrated by AGEs fluorescence, Thioflavin T (ThT) fluorescence and 1-anilinonaphthalene-8-sulphonic acid (ANS) fluorescence studies. The secondary structure of HSA was also disrupted as demonstrated by CD spectroscopy. All the parameters were nearly reverted back to native HSA formed in the glycated HSA + berberine samples. Molecular docking was utilized to identify the essential HSA residues involved in the HSA-berberine complex interaction and to ascertain the spontaneous binding of berberine to the HSA subdomain, hence favouring thermodynamic binding. The binding energy of HSA-berberine was determined to be −9.1 kcal/mol. The binding of berberine to lysine and arginine residues might be linked to its anti-glycation potential, as these amino acids play an important role in the glycation of proteins. However, further research is required to validate this assertion. Therefore, our study identifies AGEs and aggregates of the clinically significant protein HSA.

糖基化和蛋白质聚集近年来获得了更多的兴趣。糖基化导致蛋白质聚集体和晚期糖基化末端（AGEs）的形成，在几种病理条件下起着至关重要的作用。我们的研究目的是为了更深入地了解甲基乙二醛存在下AGEs的形成和人血清白蛋白（HSA）的聚集以及植物化学小檗碱的保护作用。HSA与甲基乙二醛和不同浓度的小檗碱在35-37℃下孵育7-14天。甲基乙二醛导致HSA中AGEs、纤维聚集体和疏水蛋白斑块的形成，这从AGE荧光、ThT和ANS荧光研究中可见一斑。CD光谱显示，它还破坏了HSA的二级结构。在糖化HSA +小檗碱样品中，所有这些参数都恢复到天然HSA。采用分子对接的方法鉴定了与HSA-小檗碱相互作用有关的关键HSA残基，并确定了小檗碱在HSA子结构域上的自发结合倾向于热力学结合。hsa -小檗碱的结合能为-9.1 kcal/mol。各种类型的力，如疏水相互作用、极性相互作用、氢键等，在HSA和小檗碱相互作用之间起作用。由于MGO水平在2型糖尿病等病理条件下升高，因此MGO浓度升高有可能导致HSA糖化，导致HSA水平下降，正如病理情况所观察到的那样。小檗碱与赖氨酸和精氨酸残基的结合可能与其抗糖基化潜力有关，因为这些氨基酸在蛋白质的糖基化中起重要作用。然而，需要进一步调查以证实这一说法。因此，我们的研究表征了AGEs和临床重要蛋白HSA的聚集。

{"title":"Assessing the role of Berberine as an inhibitor of advanced glycation end products (AGEs) formation using in vitro and molecular interaction studies","authors":"Yusra Ahmad , Faisal Nabi , Sana Siddiqui , Rizwan Hasan Khan , Safia Habib , Shagufta Moin","doi":"10.1016/j.abb.2025.110292","DOIUrl":"10.1016/j.abb.2025.110292","url":null,"abstract":"<div><div>Glycation leads to the formation of protein aggregates and advanced glycation end products (AGEs) by non-enzymatic reaction. AGEs have been linked to several pathological conditions such as diabetes, cardiovascular disorders, Alzheimer's etc. Our research objective is understanding how methylglyoxal triggers AGEs and protein aggregate formation in human serum albumin (HSA) and how the phytochemical berberine protects it. Employing various biochemical and biophysical techniques, we explored how berberine alters human serum albumin's biochemical properties and structure during multiple glycation stages. HSA was incubated with methylglyoxal at varying concentrations of berberine for 7–14 days at a temperature range of 35–37 degrees C. Methylglyoxal induced the formation of AGEs, fibrillar aggregates and hydrophobic protein patches in HSA as demonstrated by AGEs fluorescence, Thioflavin T (ThT) fluorescence and 1-anilinonaphthalene-8-sulphonic acid (ANS) fluorescence studies. The secondary structure of HSA was also disrupted as demonstrated by CD spectroscopy. All the parameters were nearly reverted back to native HSA formed in the glycated HSA + berberine samples. Molecular docking was utilized to identify the essential HSA residues involved in the HSA-berberine complex interaction and to ascertain the spontaneous binding of berberine to the HSA subdomain, hence favouring thermodynamic binding. The binding energy of HSA-berberine was determined to be −9.1 kcal/mol. The binding of berberine to lysine and arginine residues might be linked to its anti-glycation potential, as these amino acids play an important role in the glycation of proteins. However, further research is required to validate this assertion. Therefore, our study identifies AGEs and aggregates of the clinically significant protein HSA.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"766 ","pages":"Article 110292"},"PeriodicalIF":3.8,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142963675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Beneficial effects of Akkermansia muciniphila on benign prostatic hyperplasia and metabolic syndrome. 嗜粘阿克曼氏菌对良性前列腺增生和代谢综合征的有益作用。

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics

Pub Date : 2025-01-08 DOI: 10.1016/j.abb.2025.110294

Shengyun Wu, Xianlai Yin, Peng Yang, Binghao Gong, Zhenting Wang

Benign prostatic hyperplasia (BPH) is a prevalent condition associated with male lower urinary tract symptoms (LUTS) and is influenced by metabolic syndrome (MetS) and gut microbiota. Akkermansia muciniphila (AKK) is a gut commensal that has emerged as a potential modulator of metabolic health and inflammatory conditions. This study investigated the correlation between Akkermansia abundance and BPH severity and metabolic indices in fecal and serum samples from BPH patients and healthy donors using 16S rRNA sequencing and metabolic profiling. A testosterone-induced BPH mouse model was used to evaluate the effects of AKK administration on BPH severity and metabolic indices. Altered gut microbiota diversity was observed in BPH patients, with a significant reduction in Akkermansia abundance. Akkermansia abundance was negatively correlated with BPH symptom score, serum lipopolysaccharides (LPS), body mass index (BMI), blood glucose, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). AKK administration in BPH mice resulted in histopathological improvements, reduced prostate index, and amelioration of glandular hyperplasia. Although changes in blood glucose, TC, and LDL-C levels post-AKK supplementation were not statistically significant, a trend toward improvement was noted. Additionally, AKK administration led to a reduction in systemic inflammation markers and restoration of intestinal barrier integrity. In conclusion, AKK might modulate the gut microbiota-prostate axis and MetS. AKK's influence on systemic inflammation and gut barrier function suggests its therapeutic promise in managing BPH and associated metabolic disorders. These findings pave the way for novel microbiota-targeted therapies in the treatment of BPH and MetS.

良性前列腺增生（BPH）是一种与男性下尿路症状（LUTS）相关的普遍疾病，受代谢综合征（MetS）和肠道微生物群的影响。Akkermansia muciniphila （AKK）是一种肠道共生体，已成为代谢健康和炎症状况的潜在调节剂。本研究利用16S rRNA测序和代谢谱分析研究了Akkermansia丰度与BPH患者和健康供者粪便和血清样本中BPH严重程度和代谢指标的相关性。采用睾酮诱导的BPH小鼠模型，评价AKK给药对BPH严重程度和代谢指标的影响。在BPH患者中观察到肠道微生物群多样性的改变，Akkermansia丰度显著降低。Akkermansia丰度与BPH症状评分、血清脂多糖（LPS）、体重指数（BMI）、血糖、总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）呈负相关。在BPH小鼠中给予AKK可改善组织病理学，降低前列腺指数，改善腺体增生。虽然补充akk后血糖、TC和LDL-C水平的变化没有统计学意义，但有改善的趋势。此外，AKK管理导致全身性炎症标志物的减少和肠屏障完整性的恢复。综上所述，AKK可能调节肠道微生物群-前列腺轴和代谢代谢。AKK对全身炎症和肠道屏障功能的影响表明其在治疗BPH和相关代谢紊乱方面的治疗前景。这些发现为BPH和MetS的新型微生物靶向治疗铺平了道路。

{"title":"Beneficial effects of Akkermansia muciniphila on benign prostatic hyperplasia and metabolic syndrome.","authors":"Shengyun Wu, Xianlai Yin, Peng Yang, Binghao Gong, Zhenting Wang","doi":"10.1016/j.abb.2025.110294","DOIUrl":"https://doi.org/10.1016/j.abb.2025.110294","url":null,"abstract":"<p><p>Benign prostatic hyperplasia (BPH) is a prevalent condition associated with male lower urinary tract symptoms (LUTS) and is influenced by metabolic syndrome (MetS) and gut microbiota. Akkermansia muciniphila (AKK) is a gut commensal that has emerged as a potential modulator of metabolic health and inflammatory conditions. This study investigated the correlation between Akkermansia abundance and BPH severity and metabolic indices in fecal and serum samples from BPH patients and healthy donors using 16S rRNA sequencing and metabolic profiling. A testosterone-induced BPH mouse model was used to evaluate the effects of AKK administration on BPH severity and metabolic indices. Altered gut microbiota diversity was observed in BPH patients, with a significant reduction in Akkermansia abundance. Akkermansia abundance was negatively correlated with BPH symptom score, serum lipopolysaccharides (LPS), body mass index (BMI), blood glucose, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). AKK administration in BPH mice resulted in histopathological improvements, reduced prostate index, and amelioration of glandular hyperplasia. Although changes in blood glucose, TC, and LDL-C levels post-AKK supplementation were not statistically significant, a trend toward improvement was noted. Additionally, AKK administration led to a reduction in systemic inflammation markers and restoration of intestinal barrier integrity. In conclusion, AKK might modulate the gut microbiota-prostate axis and MetS. AKK's influence on systemic inflammation and gut barrier function suggests its therapeutic promise in managing BPH and associated metabolic disorders. These findings pave the way for novel microbiota-targeted therapies in the treatment of BPH and MetS.</p>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":" ","pages":"110294"},"PeriodicalIF":3.8,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142963676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0