Journal of Biological Chemistry最新文献_第4页

PLK1-mediated PDHA1 phosphorylation drives mitochondrial dysfunction, mitophagy, and cancer progression in Cr(VI)-associated lung cancer plk1介导的PDHA1磷酸化驱动Cr（VI）相关肺癌的线粒体功能障碍、线粒体自噬和癌症进展

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-20 DOI: 10.1016/j.jbc.2025.110406

Qiongsi Zhang, Jia Peng, Zhiguo Li, Xiongjian Rao, Derek B. Allison, Qi Qiao, Zhuangzhuang Zhang, Yifan Kong, Yanquan Zhang, Ruixin Wang, Jinghui Liu, Xinyi Wang, Chaohao Li, Fengyi Mao, Qing Shao, Tianyan Gao, Xiaoqi Liu

{"title":"PLK1-mediated PDHA1 phosphorylation drives mitochondrial dysfunction, mitophagy, and cancer progression in Cr(VI)-associated lung cancer","authors":"Qiongsi Zhang, Jia Peng, Zhiguo Li, Xiongjian Rao, Derek B. Allison, Qi Qiao, Zhuangzhuang Zhang, Yifan Kong, Yanquan Zhang, Ruixin Wang, Jinghui Liu, Xinyi Wang, Chaohao Li, Fengyi Mao, Qing Shao, Tianyan Gao, Xiaoqi Liu","doi":"10.1016/j.jbc.2025.110406","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110406","url":null,"abstract":"Hexavalent chromium (Cr(VI)) is a class I environmental carcinogen that induces lung epithelial cell transformation and promotes lung cancer progression by altering cell cycle regulation and cellular energy metabolism. In this study, we investigated the role of polo-like kinase 1 (PLK1) in Cr(VI)-transformed (CrT) bronchial epithelial cells (BEAS-2B) and found that PLK1 expression was significantly upregulated in these cells, leading to impaired mitochondrial function and enhanced mitophagy, which in turn stimulated cell proliferation both in vitro and in vivo. Mechanistically, we demonstrated that PLK1 directly phosphorylates the pyruvate dehydrogenase E1 subunit alpha 1 (PDHA1) at Thr57, leading to its destabilization and disruption of pyruvate dehydrogenase complex (PDHc) integrity. This modification inhibits oxidative phosphorylation (OXPHOS) and induces mitochondrial dysfunction. Furthermore, mitochondrial dysfunction triggers mitophagy and accelerates PDHA1 degradation, establishing a positive feedback loop that amplifies mitochondrial impairment and mitophagy, ultimately promoting cancer cell proliferation. These findings underscore the pivotal role of PLK1 in Cr(VI)-associated cancer progression and offer new insights into potential therapeutic targets to inhibit Cr(VI)-induced tumorigenesis","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"25 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144341318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Functional Genomics and Structural Insights into Maize Aldo-Keto Reductase-4 Family: Stress Metabolism and Substrate Specificity in Embryos 玉米醛酮还原酶-4家族的功能基因组学和结构分析：胚胎的应激代谢和底物特异性

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-20 DOI: 10.1016/j.jbc.2025.110404

Sylvia Morais de Sousa, Priscila Oliveira de Giuseppe, Mario Tyago Murakami, Jiahn-Chou Guan, Jonathan W. Saunders, Eduardo Kiyota, Marcelo Leite Santos, Eric A. Schmelz, Jose Andres Yunes, Karen E. Koch

{"title":"Functional Genomics and Structural Insights into Maize Aldo-Keto Reductase-4 Family: Stress Metabolism and Substrate Specificity in Embryos","authors":"Sylvia Morais de Sousa, Priscila Oliveira de Giuseppe, Mario Tyago Murakami, Jiahn-Chou Guan, Jonathan W. Saunders, Eduardo Kiyota, Marcelo Leite Santos, Eric A. Schmelz, Jose Andres Yunes, Karen E. Koch","doi":"10.1016/j.jbc.2025.110404","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110404","url":null,"abstract":"Aldo-keto reductases (AKRs) are ubiquitous in nature and are able to reduce a wide range of substrates, from simple sugars to potentially toxic aldehydes. In plants, AKRs are involved in key metabolic processes including reactive aldehyde detoxification. This study aimed to i) delineate a maize gene family encoding Aldo Keto Reductase-4s (AKR4s) ii) help bridge sequence-to-function gaps among them, and iii) focus on a family member implicated in embryo specific stress metabolism. We employed a genome-wide analysis approach to identify maize genes encoding AKR4s, defining and annotating a 15-member gene family that clustered into three subgroups. Expression profiling, validated through wet lab experiments, revealed distinct functional roles: i) AKR4C Zm-1 functions in aldehyde detoxification during stress, ii) AKR4C Zm-2 includes stress-responsive AKRs with diverse substrate affinities, and iii) AKR4A/B Zm-3 contributes to specialized metabolites like phytosiderophores for iron transport. To investigate the impact of sequence variation on function, we characterized ZmAKR4C13, a representative of AKR4C Zm-1. Its mRNA and protein were predominantly localized in embryos, suggesting a specialized role. Recombinant ZmAKR4C13 efficiently reduced methylglyoxal and small aldehydes but showed poor activity toward aldoses larger than four carbons. Crystallographic analysis identified a size constraint at the active site, attributed to the bulkier LEU residue at position 294. Collectively, our results emphasize how subtle modifications in active-site architecture influence AKR substrate specificity. They also demonstrate a potential role of maize ZmAKR4C13 in detoxifying methylglyoxal and other small metabolites that could contribute to stress signaling in embryos.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"15 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144341320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

HSD3B1 Upregulation via LRH1 Sustains Estrogen Receptor Signaling and Promotes Endocrine Resistance in Breast Cancer 通过LRH1上调HSD3B1维持雌激素受体信号传导并促进乳腺癌内分泌抵抗

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-20 DOI: 10.1016/j.jbc.2025.110405

Xiuxiu Li, Yoon-Mi Chung, Monaben Patel, Nima Sharifi

{"title":"HSD3B1 Upregulation via LRH1 Sustains Estrogen Receptor Signaling and Promotes Endocrine Resistance in Breast Cancer","authors":"Xiuxiu Li, Yoon-Mi Chung, Monaben Patel, Nima Sharifi","doi":"10.1016/j.jbc.2025.110405","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110405","url":null,"abstract":"Endocrine resistance is a major challenge in the treatment of estrogen receptor-positive (ER<ce:sup loc=\"post\">+</ce:sup>) breast cancer, often leading to disease recurrence and metastasis. 3β-Hydroxysteroid dehydrogenase 1 (3βHSD1, encoded by <ce:italic>HSD3B1</ce:italic>) catalyzes the rate-limiting conversion of dehydroepiandrosterone (DHEA) to androstenedione (AD), the major substrate for aromatase and a key precursor for estrogen biosynthesis. However, the regulation of <ce:italic>HSD3B1</ce:italic> in endocrine-resistant breast cancer remains unclear. We show that long-term estrogen deprivation (LTED) or tamoxifen treatment induces <ce:italic>HSD3B1</ce:italic> expression and enzymatic activity, sustaining DHEA metabolism and ER signaling in resistant ER<ce:sup loc=\"post\">+</ce:sup> breast cancer cells. T47D-LTED and T47D-4OHT cells exhibited increased <ce:italic>HSD3B1</ce:italic> expression and enhanced DHEA metabolism. <ce:italic>HSD3B1</ce:italic> deficiency impaired DHEA-driven survival, confirming its role in endocrine resistance. Mechanistically, we identify liver receptor homolog-1 (LRH1/NR5A2) as a key transcriptional regulator of <ce:italic>HSD3B1</ce:italic>. LRH1 inhibition suppressed <ce:italic>HSD3B1</ce:italic> expression, DHEA metabolism, and ER target gene activation, demonstrating its role in sustaining estrogen synthesis and tumor adaptation. Our findings establish <ce:italic>HSD3B1</ce:italic> as a critical mediator of endocrine resistance and identify LRH1 as an upstream regulator. Targeting <ce:italic>HSD3B1</ce:italic> or LRH1 may offer a new therapeutic strategy to restore endocrine sensitivity in ER<ce:sup loc=\"post\">+</ce:sup> breast cancer.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"7 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144341319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

A tubulin-binding protein that preferentially binds to GDP-tubulin and promotes GTP exchange. 一种优先结合gdp -微管蛋白并促进GTP交换的微管蛋白结合蛋白。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110401

Wesley J Yon, Taekjip Ha, Yixian Zheng, Ross T A Pedersen

引用次数: 0

CryoEM Structure of Rv2531c Reveals Cofactor-Induced Tetramer-Dimer Transition in a Tuberculin Amino Acid Decarboxylase. Rv2531c的低温结构揭示了辅助因子在结核菌素氨基酸脱羧酶中诱导的四聚体-二聚体转变。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110394

Jyoti Gupta,Tina Izard

{"title":"CryoEM Structure of Rv2531c Reveals Cofactor-Induced Tetramer-Dimer Transition in a Tuberculin Amino Acid Decarboxylase.","authors":"Jyoti Gupta,Tina Izard","doi":"10.1016/j.jbc.2025.110394","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110394","url":null,"abstract":"The survival of Mycobacterium tuberculosis relies on its ability to adapt to dynamic and hostile host environments. Amino acid decarboxylases play a crucial role in these adaptations, but their structural and mechanistic properties are not fully understood. Bioinformatic analyses revealed that these enzymes exist in three distinct forms based on their domain organization. We used cryogenic electron microscopy (cryoEM) at 2.86 Å resolution to show that Rv2531c exhibits unexpected oligomeric and conformational flexibility. The enzyme forms a tetramer with distinct open and closed conformations in its apo state, suggesting dynamic inter-subunit interactions. Upon binding pyridoxal 5'-phosphate (PLP), the enzyme undergoes a dramatic structural rearrangement, transitioning into a dimer. These findings reveal a novel mechanism of oligomeric plasticity. We also uncover an amino-terminal domain that might play a role in this process. Our results provide critical insights into the structural adaptations that support bacterial persistence under intracellular stress. By elucidating the apo and PLP-bound states of Rv2531c, we contribute to a deeper understanding of how M. tuberculosis navigates its challenging intracellular environment. These insights into the unique structural features of Rv2531c offer a foundation for targeting metabolic resilience in tuberculosis and open avenues for future studies on the role of this domain in pathogenesis.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"56 1","pages":"110394"},"PeriodicalIF":4.8,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337460","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Rag GTPases control lysosomal acidification by regulating v-ATPase assembly in Drosophila. Rag gtpase通过调节果蝇v- atp酶的组装来控制溶酶体酸化。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110400

Ying Zhou,Xiaodie Yang,Wenyu Xu,Sulin Shen,Weikang Fan,Guoqiang Meng,Yang Cheng,Yingying Lu,Youheng Wei

{"title":"Rag GTPases control lysosomal acidification by regulating v-ATPase assembly in Drosophila.","authors":"Ying Zhou,Xiaodie Yang,Wenyu Xu,Sulin Shen,Weikang Fan,Guoqiang Meng,Yang Cheng,Yingying Lu,Youheng Wei","doi":"10.1016/j.jbc.2025.110400","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110400","url":null,"abstract":"The Rag GTPases play an important role in sensing amino acids and activating the target of rapamycin complex 1 (TORC1), a master regulator of cell metabolism. Previously, we have shown that GDP-bound RagA stimulates lysosome acidification and autophagic degradation, which are essential for young egg chamber survival under starvation in Drosophila. However, the underlying mechanism is unclear. Here we demonstrate that the GDP-bound RagA breaks the physical interaction between chaperonin containing tailless complex polypeptide 1 (CCT) and Vacuolar H+-ATPase (v-ATPase) subunit V1, and thus promotes the assembly of active v-ATPase and increases the lysosomal acidification. Consistently, knockdown of CCT complex components rescued the accumulation of defective autolysosomes in RagA RNAi. Moreover, the knockdown of Lamtor4, a component of lysosomal adaptor and MAPK and mTOR activator (LAMTOR) that anchors Rag GTPases to the lysosome, resulted in autolysosome accumulation, suggesting that RagGTPases regulate lysosomal acidification depend on their lysosomal localization. Knockdown of the CCT complex components attenuated the autophagic defects in Lamtor 4 RNAi. Our work highlights the interaction between CCT and v-ATPase in regulating lysosomal acidification.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"70 1","pages":"110400"},"PeriodicalIF":4.8,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

5hmC enhances PARP trapping and restores PARP inhibitor sensitivity in chemoresistant BRCA1/2-deficient cells. 5hmC在耐药brca1 /2缺陷细胞中增强PARP捕获并恢复PARP抑制剂敏感性。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110393

Suhas S Kharat, Arun P Mishra, Satheesh K Sengodan, Dillon Dierman, Stephen D Fox, Walter J Chazin, Shyam K Sharan

引用次数: 0

How short is too short for amyloid fibrils?: molecular dynamics of oligomers of infectious prion core structures. 淀粉样蛋白原纤维有多短才算太短？感染性朊病毒核心结构低聚物的分子动力学。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110390

Efrosini Artikis,Amitava Roy,Byron Caughey

{"title":"How short is too short for amyloid fibrils?: molecular dynamics of oligomers of infectious prion core structures.","authors":"Efrosini Artikis,Amitava Roy,Byron Caughey","doi":"10.1016/j.jbc.2025.110390","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110390","url":null,"abstract":"In many proteinopathies, the relative conformations of amyloid fibrils versus smaller oligomers remain unclear. Most tissue-derived isolates of infectious prion protein (PrP) prions are predominantly fibrillar. A few studies have asserted that prion amyloid fibrils efficiently disassemble into dimeric to tetrameric \"elemental bricks\" under certain detergent or chaotropic conditions, but our companion paper provides strong evidence to the contrary. Given our difficulties in isolating detectable amounts of small oligomeric (2-4-mer) prions, we performed molecular dynamics simulations to test the abilities of small fragments (dimers to 25-mers) of cryo-EM-based infectious prion fibril core structures to retain their conformational integrity. We show that dimers of the aRML prion structure lost most of their original secondary and tertiary structure within <<1 μs, while trimers maintained some intermolecular β-sheets. Further increases in fragment size helped preserve major structural motifs and the integrity of the templating surfaces responsible for self-propagation. In simulations of octamers and/or 25-mers, even at elevated temperatures, no fragmentation was observed for either the aRML, 22L, or 263K prion strains, although the terminal chains were substantially destabilized. Together, our results provide evidence that oligomeric fragments of prion fibril cores as small as tetramers retain substantial structural integrity. Our findings suggest that, as exemplified by PrP fibrils, short cores as small as tetramers may be stable enough to account for bioactive oligomeric species detected in brain extracts from proteinopathy patients. However, the lack of observed spontaneous core fragmentation suggests that prion oligomers might be rare in vivo and/or producedby non-autonomous physiological cleavage processes.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"25 1","pages":"110390"},"PeriodicalIF":4.8,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Crosstalk between iron and flavins in the opportunistic fungal pathogen Candida albicans. 机会真菌病原体白色念珠菌中铁与黄素的串扰。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110396

Marika S David,Zhengkai Zhu,Maranda R McDonald,Mohsen Badiee,I Phillip Mortimer,Anthony K L Leung,Valeria C Culotta

{"title":"Crosstalk between iron and flavins in the opportunistic fungal pathogen Candida albicans.","authors":"Marika S David,Zhengkai Zhu,Maranda R McDonald,Mohsen Badiee,I Phillip Mortimer,Anthony K L Leung,Valeria C Culotta","doi":"10.1016/j.jbc.2025.110396","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110396","url":null,"abstract":"As part of the innate immune response, the host withholds metal micronutrients such as iron (Fe) from invading pathogens. To survive such Fe-limitation, the opportunistic fungal pathogen Candida albicans has evolved elaborate responses to Fe-starvation stress. One such adaptation involves secretion of flavins, yellow isoalloxazine compounds that serve important redox roles in biology. Why the organism secretes flavins during Fe-starvation is not known. Moreover, the exact flavin secreted by the fungus or the effects of Fe-starvation on intracellular flavin pools has not been documented. Major cellular flavins include riboflavin (vitamin B2) and the downstream metabolites and enzyme co-factors FAD and FMN. Of these, our HPLC and mass spectrometry analyses identify riboflavin as the sole flavin secreted by Fe-starved C. albicans. Fe also regulates intracellular pools of flavins. While Fe-replete cells have abundant FMN and FAD with only trace riboflavin, Fe-starvation induces a spike in intracellular riboflavin, whereas FMN and FAD are unaffected. These shifts in riboflavin are dependent on the Fe-sensing transcription factor Sef1. During Fe-starvation, Sef1 induces genes for riboflavin biosynthesis, but not for the conversion of riboflavin to FMN and FAD. Sef1 is also needed to activate riboflavin export. We provide evidence for the first time that extracellular riboflavin can enhance fungal uptake of Fe. Specifically, riboflavin increased C. albicans acquisition of Fe from animal serum, presumably through its redox activity on extracellular Fe. Our observed role of riboflavin on Fe uptake may promote C. albicans fitness in the Fe-limiting environment of the host.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"1 1","pages":"110396"},"PeriodicalIF":4.8,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Unique structural features define the decarboxylation activity of a CYP152 fatty acid decarboxylase from Lacicoccus alkaliphilus. 嗜碱Lacicoccus alkaliphilus独特的结构特征确定了CYP152脂肪酸脱羧酶的脱羧活性。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-19 DOI: 10.1016/j.jbc.2025.110397

Suppalak Phaisan,Aisaraphon Phintha,Duangthip Trisrivirat,Narin Lawan,Jeerus Sucharitakul,Ailada Charoenpol,Pratchaya Watthaisong,Hideaki Tanaka,Genji Kurisu,Pimchai Chaiyen

{"title":"Unique structural features define the decarboxylation activity of a CYP152 fatty acid decarboxylase from Lacicoccus alkaliphilus.","authors":"Suppalak Phaisan,Aisaraphon Phintha,Duangthip Trisrivirat,Narin Lawan,Jeerus Sucharitakul,Ailada Charoenpol,Pratchaya Watthaisong,Hideaki Tanaka,Genji Kurisu,Pimchai Chaiyen","doi":"10.1016/j.jbc.2025.110397","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110397","url":null,"abstract":"Cytochrome P450 CYP152s catalyze decarboxylation of fatty acids to generate terminal alkenes, valuable compounds for various industries. Here, we identified, overexpressed, and characterized a new CYP152 enzyme from Lacicoccus alkaliphilus (OleTLA), and compared its biophysical and biochemical properties with the well-studied OleTJE from Jeotgalicoccus sp. 8456. Improved expression protocols gave the highest yields of CYP152 holoenzymes reported to date. OleTLA exhibits twice the catalytic turnover number of OleTJE when using hexadecanoic acid and H2O2 as substrates in 10% (v/v) ethanol. The X-ray structure of OleTLA in complex with icosanoic acid revealed a unique flipped heme and a substrate tunnel configuration which are different than those of other CYP152 decarboxylases. Molecular dynamics simulations revealed that in the presence of EtOH, OleTLA displays structural dynamics which maintain structural interactions better than those of OleTJE. As I178 in OleTLA (equivalent to L176 in OleTJE) shows close interactions with its substrate during simulations, I178L of OleTLA and L176I of OleTJE variants were constructed and investigated for their activities. While L176I in OleTJE caused a significant loss of activity, I178L of OleTLA had activities that were equivalent to or greater than those of the wild-type enzyme, suggesting that overall scaffold of OleTLA is more amenable to mutation than OleTJE. Stopped-flow investigations of OleTLA reactions indicated that EtOH increases the rate constant of Compound I formation. We also identified a new redox partner system, ferredoxin and ferredoxin reductase that can function as effective electron donors for both in vitro and in vivo systems of CYP152s.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"45 1","pages":"110397"},"PeriodicalIF":4.8,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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