Journal of Biological Chemistry最新文献

YEATS2 O-GlcNAcylation Promotes Chromatin Association of the ATAC Complex and Lung Cancer Tumorigenesis. YEATS2 o - glcn酰化促进ATAC复合物的染色质关联和肺癌的发生。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-18 DOI: 10.1016/j.jbc.2025.110388

Jianxin Zhao,Zheng Zhao,Wen Zhou,Jianzhi Zhang,Jinfeng Chen,Jianwei Sun,Jing Li

{"title":"YEATS2 O-GlcNAcylation Promotes Chromatin Association of the ATAC Complex and Lung Cancer Tumorigenesis.","authors":"Jianxin Zhao,Zheng Zhao,Wen Zhou,Jianzhi Zhang,Jinfeng Chen,Jianwei Sun,Jing Li","doi":"10.1016/j.jbc.2025.110388","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110388","url":null,"abstract":"The intracellular O-linked N-acetylglucosamine (O-GlcNAc) modification is known to be enriched in the nucleus and on chromatin, but many of its chromatin targets remain to be identified. Herein we demonstrate the O-GlcNAcylation of YEATS Domain Containing 2 (YEATS2), a subunit of the chromatin Ada-two-A-containing (ATAC) complex and a reader of histone H3K27ac. We show that YEATS2 interacts with the O-GlcNAc transferase (OGT) and further pinpoint its major O-GlcNAcylation site to be Thr604 using electron transfer dissociation mass spectrometry. O-GlcNAcylation promotes the chromatin association of YEATS2, and the affinity between YEATS2 and other ATAC components on chromatin, such as ZZZ3, GCN5 and PCAF. Downstream, YEATS2-T604A mutants attenuated the ATAC-dependent histone H3K9ac levels and inactivated the expression of essential ribosomal genes as shown in chromatin immunoprecipitation assays. Further, xenograft experiments show that YEATS2 O-GlcNAcylation promotes lung cancer tumorigenesis. Our work reveals the critical role of YEATS2 O-GlcNAcylation in stabilizing the ATAC complex on chromatin and expands the chromatin substrates of OGT.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"179 1","pages":"110388"},"PeriodicalIF":4.8,"publicationDate":"2025-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144335305","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

TFAP2C Drives Cisplatin Resistance in Bladder Cancer by Upregulating YAP and Activating β-Catenin Signaling. TFAP2C通过上调YAP和激活β-Catenin信号通路驱动膀胱癌顺铂耐药

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-18 DOI: 10.1016/j.jbc.2025.110387

Qiufeng Pan,Changmin Zou,Zhigen Lin,Hao Tang,Zepu Long,Longwang Wang

{"title":"TFAP2C Drives Cisplatin Resistance in Bladder Cancer by Upregulating YAP and Activating β-Catenin Signaling.","authors":"Qiufeng Pan,Changmin Zou,Zhigen Lin,Hao Tang,Zepu Long,Longwang Wang","doi":"10.1016/j.jbc.2025.110387","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110387","url":null,"abstract":"Cisplatin-based chemotherapy is a conventional therapy for muscle-invasive bladder cancer (BC); However, its efficacy is often limited by the emergence of resistance to cisplatin. Yes-associated protein (YAP) and β-catenin are involved in this resistance, yet their upstream regulators are not well defined. This study investigates the role of TFAP2C in regulating YAP expression and its impact on cisplatin resistance in BC. The Cancer Genome Atlas (TCGA) gene expression data and GSE231835 dataset were analyzed to identify potential transcription factors regulating YAP. Assessed TFAP2C and YAP expression in clinical samples and cell lines. Functional assays were performed following TFAP2C knockdown. Dual-luciferase reporter assays and Chromatin immunoprecipitation (ChIP) confirmed TFAP2C binding to the YAP promoter. An mouse model evaluated the effects of TFAP2C silencing on tumor growth and cisplatin resistance. The results showed that TFAP2C was identified as an upstream activator of YAP, with elevated expression in cisplatin-resistant BC cell lines and positive correlation with YAP expression. Silencing TFAP2C reduced malignant behaviors, decreased YAP, phosphorylated YAP (p-YAP), and β-catenin levels, and increased apoptosis in both cisplatin-sensitive and resistant BC cells. Besides, TFAP2C directly binds to the YAP promoter, enhancing its transcription. In the xenograft model, TFAP2C silencing significantly inhibited tumor growth and reduced cisplatin resistance. TFAP2C promotes cisplatin resistance and malignant behavior in BC by upregulating YAP and activating the β-catenin signaling pathway. Targeting TFAP2C offers a novel therapeutic strategy to overcome cisplatin resistance in BC, representing a new discovery in combating chemoresistance.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"24 1","pages":"110387"},"PeriodicalIF":4.8,"publicationDate":"2025-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144335359","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

The Escherichia coli RelB antitoxin C-terminus is essential for RelE toxin suppression and transcriptional repression. 大肠杆菌RelB抗毒素c端对RelB毒素抑制和转录抑制至关重要。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-18 DOI: 10.1016/j.jbc.2025.110389

Julia Tanquary,Ian J Pavelich,Marcin Grabowicz,Christine M Dunham

{"title":"The Escherichia coli RelB antitoxin C-terminus is essential for RelE toxin suppression and transcriptional repression.","authors":"Julia Tanquary,Ian J Pavelich,Marcin Grabowicz,Christine M Dunham","doi":"10.1016/j.jbc.2025.110389","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110389","url":null,"abstract":"Bacterial type II toxin-antitoxin (TA) systems exhibit high specificity within each pair to ensure precise recognition of the toxin by its cognate antitoxin to inhibit toxicity of the free toxin. Despite high structural similarity among some TAs, crosstalk between non-cognate TA pairs is rare. To determine how the E. coli RelB antitoxin suppresses its cognate RelE toxin, we engineered C-terminal truncations of RelB and tested their functional effects on toxin suppression in E. coli. We find that removal of the long C-terminal α3 and connecting loop 4 (L4) of RelB prevents RelE suppression. Quantitative binding assays of RelE and RelB variants support a reduced affinity upon removal of the RelB C-terminus. Disrupting these interactions between RelB and RelE also led to a significant decrease in transcriptional repression at the relO operator, underscoring the requirement for RelE binding to RelB for optimal repression at DNA repressor elements. Comparison to other structurally homologous TA systems, such as E. coli DinJ-YafQ, reveals key differences in the molecular mechanisms of both toxin suppression and DNA repressor activity highlighting the diversity in TA regulation and function.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"14 1","pages":"110389"},"PeriodicalIF":4.8,"publicationDate":"2025-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144335358","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

The redox landscape of pyruvate:ferredoxin oxidoreductases reveals often conserved Fe-S cluster potentials. 丙酮酸：铁氧还蛋白氧化还原酶的氧化还原景观揭示了通常保守的Fe-S簇电位。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-16 DOI: 10.1016/j.jbc.2025.110380

Sheila C Bonitatibus,Ryan V Pham,Andrew C Weitz,Madeline M Lopéz-Muñoz,Bin Li,William W Metcalf,Satish K Nair,Sean J Elliott

{"title":"The redox landscape of pyruvate:ferredoxin oxidoreductases reveals often conserved Fe-S cluster potentials.","authors":"Sheila C Bonitatibus,Ryan V Pham,Andrew C Weitz,Madeline M Lopéz-Muñoz,Bin Li,William W Metcalf,Satish K Nair,Sean J Elliott","doi":"10.1016/j.jbc.2025.110380","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110380","url":null,"abstract":"Here we investigate the thermodynamic driving force of internal electron transfer (ET) of pyruvate:ferredoxin oxidoreductases (PFORs), by comparing the redox properties of a series of PFORs from Chlorobaculum tepidum (Ct), Magnetococcus marinus (Mm), Methanosarccina acetivorans (Ma), as well as revisiting the single historical precedent, the enzyme from Desulfovibrio africanus. These enzymes require a thiamine pyrophosphate (TPP) cofactor, three [4Fe-4S] clusters, and Coenzyme A (CoA) for activity and are found within anaerobic organisms that utilize the reverse tricarboxylic acid (TCA cycle), or other reductive pathways, performing CO2 reduction and pyruvate synthesis. Yet, PFOR is often invoked as an oxidative enzyme responsible for generating reducing equivalents in the form of the redox carrier ferredoxin. Previous efforts to understand the mechanism of PFOR have relied upon a prior report of the iron-sulfur redox potentials derived from an incomplete redox titration. Here we use direct protein film electrochemistry (PFE) to provide a side-by-comparison of four PFOR enzymes, providing a new assessment of the iron-sulfur cluster redox potentials. As the Ma PFOR is comprised of multiple polypeptides, our investigation of the recombinant PorD sub-unit allows us to construct a model where the revised redox-potentials are mapped to specific iron-sulfur clusters.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"145 1","pages":"110380"},"PeriodicalIF":4.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144320235","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

CpgD is a phosphoglycerate cytidylyltransferase required for ceramide diphosphoglycerate synthesis CpgD是合成二磷酸甘油酸神经酰胺所需的磷酸甘油酸胞基转移酶

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-16 DOI: 10.1016/j.jbc.2025.110386

Tanisha Dhakephalkar, Ziqiang Guan, Eric A. Klein

{"title":"CpgD is a phosphoglycerate cytidylyltransferase required for ceramide diphosphoglycerate synthesis","authors":"Tanisha Dhakephalkar, Ziqiang Guan, Eric A. Klein","doi":"10.1016/j.jbc.2025.110386","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110386","url":null,"abstract":"LPS is essential in most Gram-negative bacteria, but mutants of several species have been isolated that can survive in its absence. <ce:italic>Caulobacter crescentus</ce:italic> viability in the absence of LPS is partially dependent on the anionic sphingolipid ceramide diphosphoglycerate (CPG2). Genetic analyses showed that <ce:italic>ccna_01210</ce:italic>, which encodes a nucleotidyltransferase, is required for CPG2 production. Using purified recombinant protein, we determined that CCNA_01210 (CpgD) is a phosphoglycerate cytidylyltransferase which uses CTP and phosphoglycerate to produce CDP-glycerate, which we hypothesize is the phosphoglycerate donor for CPG2 synthesis. CpgD had optimum activity at pH 7.5-8 in the presence of magnesium. CpgD exhibited Michaelis-Menten kinetics with respect to 3-phosphoglycerate, D-2-phosphoglycerate, and L-2-phosphoglycerate. By contrast, CTP followed Michaelis-Menten kinetics in the presence of 3-phosphoglycerate and L-2-phosphosglycerate but exhibited cooperativity with D-2-phosphoglycerate. Overall, D-2-phosphoglycerate was the preferred substrate <ce:italic>in vitro</ce:italic>. The characterization of this enzyme uncovers another step in the pathway towards CPG2 synthesis.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"6 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144305069","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 kinetic model of copper homeostasis in Saccharomyces cerevisiae. 酿酒酵母菌铜稳态的动力学模型。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-16 DOI: 10.1016/j.jbc.2025.110368

Cade Dulaney,Jay R Walton,Paul A Lindahl

{"title":"A kinetic model of copper homeostasis in Saccharomyces cerevisiae.","authors":"Cade Dulaney,Jay R Walton,Paul A Lindahl","doi":"10.1016/j.jbc.2025.110368","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110368","url":null,"abstract":"Rather than inhibiting copper entry when grown on high Cu, yeast cells import excessive Cu while simultaneously increasing expression of metallothionein CUP1 which then sequesters the excess Cu. An ordinary-differential-equations-based kinetic model was developed to investigate this unusual behavior. The assumed reaction network included 25 reactions and 10 components in the cytosol of yeast cells growing in media supplemented with increasing nutrient COPPER concentrations. Published concentrations of copper proteins and coordination complexes that constitutes the low-molecular-mass (or labile) Cu pool were assumed. Other components included transcription factors MAC1 and ACE1, the MAC1-dependent copper importer CTR1, and other copper proteins considered collectively. A second MAC1-independent importer was required for sufficient Cu to enter the cell under Cu-excess conditions. The mathematical system was initially solved at steady-state for each condition in the series. The null-space of the stoichiometric matrix was evaluated using the Basic Pathways approach. Steady-state rates and rate-constants were calculated for each reaction and each condition of the series. Four rate-constants trended higher across the series indicating that the cell regulates those reactions in ways that were not included in the assumed rate-law expressions. This behavior was simulated by augmenting those expressions with logistical functions that sensed labile Cu and/or nutrient COPPER. The resulting integrated dynamical system approximately generated observed component concentrations over the series and was stable to both intracellular and extracellular perturbations. The MAC1-independent importer is predicted to be FET4, a nonspecific importer of both Cu and Fe. Cells likely tolerate excessive Cu import to import sufficient iron.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"101 1","pages":"110368"},"PeriodicalIF":4.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144320323","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

Molecular arrangements that accompany binding of rice xylanase inhibitor protein OsXIP and the Rhizopus oryzae GH11 xylanase RXyn2. 水稻木聚糖酶抑制蛋白OsXIP与米根霉GH11木聚糖酶RXyn2结合的分子排列

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-16 DOI: 10.1016/j.jbc.2025.110385

Takayuki Ohnuma,Jun Tanaka,Harutada Ozaki,Keigo Mitsui,Daichi Tsujitsugu,Miki Okugawa,Toru Takeda,Makoto Ihara,Tamo Fukamizo,Daijiro Takeshita

{"title":"Molecular arrangements that accompany binding of rice xylanase inhibitor protein OsXIP and the Rhizopus oryzae GH11 xylanase RXyn2.","authors":"Takayuki Ohnuma,Jun Tanaka,Harutada Ozaki,Keigo Mitsui,Daichi Tsujitsugu,Miki Okugawa,Toru Takeda,Makoto Ihara,Tamo Fukamizo,Daijiro Takeshita","doi":"10.1016/j.jbc.2025.110385","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110385","url":null,"abstract":"Plants have evolved xylanase inhibitor proteins as part of their defense mechanisms against phytopathogens. The rice xylanase inhibitor protein (OsXIP) is structurally similar to GH18 chitinase and homologous to wheat XIP-type inhibitor (XIP-I), which inhibits both GH10 and GH11 xylanases. Various inhibition and interaction analyses showed that OsXIP competitively inhibits the hydrolytic activity of GH11 xylanase RXyn2, but not the activity of GH10 xylanase RXyn1 from Rhizopus oryzae. The crystal structure of the OsXIP/RXyn2 complex showed that OsXIP, which has a (β/α)8-barrel fold, extrudes the loop between α4 and β5 (Lα4β5OsXIP) and inserts the loop into the xylotriose binding site (-3 to -1 subsite) formed by the inner β-sheet (palm) of RXyn2 jelly roll. The guanidyl group of Arg155 in Lα4β5OsXIP was shown to be critical for the inhibitory activity by mutational analysis. Notably, in the complex structure, the cylindrical cavity formed by the palm of RXyn2 jelly roll stacked upright on the loops at the N-terminal ends of the β-strands of OsXIP (I-formation). On the other hand, in the complex structure of XIP-I and GH11 xylanase from Talaromyces funiculosus (XYNC), the cavity of XYNC laid tangentially to the part of the corresponding region of XIP-I through the Lα4β5XIP-I (T-formation). The dissociation constant of the OsXIP/RXyn2 complex was one tenth of that of the XIP-I/XYNC complex (4.2 versus 41.5 nM). OsXIP may have adapted to bind and inhibit GH11 enzymes, which are resistant to the inhibition by XIP-I type proteins, by changing its binding mode.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"23 1","pages":"110385"},"PeriodicalIF":4.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144320324","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

Structural biology of Parkinson’s Disease-associated Leucine-Rich Repeat Kinase 2 (LRRK2) 帕金森病相关富亮氨酸重复激酶2 （LRRK2）的结构生物学研究

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-14 DOI: 10.1016/j.jbc.2025.110376

Andres E. Leschziner

引用次数: 0

FAK activity exacerbates disturbed flow-mediated atherosclerosis via VEGFR2-CBL-NF-κB signaling FAK活性通过VEGFR2-CBL-NF-κB信号加剧了紊乱的血流介导的动脉粥样硬化

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-14 DOI: 10.1016/j.jbc.2025.110383

James M. Murphy, Duyen Thi Kieu Tran, Kyuho Jeong, Ly Nguyen, Mai Thi Nguyen, Dhananjay Tambe, Hanjoong Jo, Eun-Young Erin Ahn, Ssang-Taek Steve Lim

{"title":"FAK activity exacerbates disturbed flow-mediated atherosclerosis via VEGFR2-CBL-NF-κB signaling","authors":"James M. Murphy, Duyen Thi Kieu Tran, Kyuho Jeong, Ly Nguyen, Mai Thi Nguyen, Dhananjay Tambe, Hanjoong Jo, Eun-Young Erin Ahn, Ssang-Taek Steve Lim","doi":"10.1016/j.jbc.2025.110383","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110383","url":null,"abstract":"Atherosclerosis develops at predictable sites in the vasculature where branch points and curvatures create non-laminar disturbed flow. This disturbed flow causes vascular inflammation by increased endothelial cell (EC) barrier permeability and the expression of inflammatory genes such as vascular cell adhesion molecule-1 (VCAM-1). Vascular endothelial growth factor receptor 2 (VEGFR2) regulates flow-induced EC inflammation; however, there are still some gaps in understanding the precise signaling mechanism or pathway. Focal adhesion kinase (FAK) is a protein tyrosine kinase whose expression has been implicated in flow-mediated signaling in ECs. However, the link between FAK and VEGFR2 in flow-mediated inflammation signaling has remained unelucidated. Here we found that priming of VEGFR2 with VEGF was critical for flow-mediated activation of FAK and NF-kB. Mechanistically, FAK activation triggers tyrosine phosphorylation of Casitas B-lineage lymphoma (CBL; an E3 ubiquitin ligase) that interacts with VEGFR2 under flow conditions. However, FAK inhibition reduced VEGFR2-FAK-CBL complex formation, partly due to reduced expression of VEGFR2 on the cell membrane. Further, <ce:italic>Apoe-/-</ce:italic> mice fed a western diet (WD) exhibited increased FAK activity within the atheroprone disturbed flow region of the inner aortic arch compared to the outer arch. Disturbed flow-induced FAK activation is associated with elevated VEGFR2 on the surface of ECs of the inner aortic arch, but not in the outer arch. Taken together, these data suggest that suppression of augmented FAK activity under disturbed flow may prove beneficial in reducing pro-inflammatory signaling of the endothelial layer.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"625 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144305073","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

Characterization of the activity of KTX-1001, a small molecule inhibitor of multiple myeloma SET domain (MMSET) using surface plasmon resonance (SPR) 利用表面等离子体共振（SPR）表征多发性骨髓瘤SET结构域（MMSET）小分子抑制剂KTX-1001的活性

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-06-14 DOI: 10.1016/j.jbc.2025.110382

Chad A. Lewis, Charles Schmidt, Lisa Beebe, Terrence J. Connolly

{"title":"Characterization of the activity of KTX-1001, a small molecule inhibitor of multiple myeloma SET domain (MMSET) using surface plasmon resonance (SPR)","authors":"Chad A. Lewis, Charles Schmidt, Lisa Beebe, Terrence J. Connolly","doi":"10.1016/j.jbc.2025.110382","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110382","url":null,"abstract":"KTX-1001 is a small molecule inhibitor of MMSET in early clinical development for multiple myeloma. It was identified as a potent and selective inhibitor of MMSET (also known as NSD2) using a high throughput biochemical assay with LC/MS-MS detection of SAH production as the endpoint. Subsequent evaluation of the binding of KTX-1001 to its target was conducted using surface plasmon resonance (SPR) to quantify on-rate, off-rate, and equilibrium dissociation constant utilizing the SET domain as the immobilized target. In this format, no saturable or specific binding could be observed, despite the potent activity in the assay using full length NSD2 and radiolabeled SAM. To interrogate the discordance between potent activity and the lack of detectable binding in the assay, a series of experiments were designed in which KTX-1001 with a biotin-PEG tether (KTX-1001-3) was immobilized to the chip, with target (NSD2 SET domain) in-flow, with nucleosomes and with and without cofactor. These experiments demonstrated that KTX-1001-3 bound to the SET domain in a specific and saturable manner, with an affinity comparable to the IC<ce:inf loc=\"post\">50</ce:inf> determined in the enzymatic assay. Further, these studies confirmed unique binding properties of KTX-1001 in the presence of nucleosomes, cofactor, and in combination. These data identify the utility of SPR in a “reverse” format, where immobilization of KTX-1001 allowed for the interrogation of binding to a protein target that may be challenging if conformation changes are induced in the coupling of that biologic target to a chip surface. Collectively, this analysis demonstrates the specific potent biochemical activity of KTX-1001 against MMSET and supports the ongoing evaluation of KTX-1001 in the clinic.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"142 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144305071","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