FEBS LettersPub Date : 2025-01-17DOI: 10.1002/1873-3468.15095
Lars Kaduhr, Klaus Mayer, Raffael Schaffrath, Johannes Buchner, Ulrich Brinkmann
{"title":"Diphthamide synthesis is linked to the eEF2-client chaperone machinery.","authors":"Lars Kaduhr, Klaus Mayer, Raffael Schaffrath, Johannes Buchner, Ulrich Brinkmann","doi":"10.1002/1873-3468.15095","DOIUrl":"https://doi.org/10.1002/1873-3468.15095","url":null,"abstract":"<p><p>The diphthamide modification of eukaryotic translation elongation factor (eEF2) is important for accurate protein synthesis. While the enzymes for diphthamide synthesis are known, coordination of eEF2 synthesis with the diphthamide modification to maintain only modified eEF2 is unknown. Physical and genetic interactions extracted from BioGRID show a connection between diphthamide synthesis enzymes and chaperones in yeast. This includes the Hsp90 co-chaperones Hgh1 and Cpr7. The respective co-chaperone deletion strains contained eEF2 without diphthamide. Notably, strains deficient in other co-chaperones showed no defect in the eEF2-diphthamide modification. Our results demonstrate that diphthamide synthesis involves not only Dph enzymes but also the eEF2-interacting co-chaperones Hgh1 and Cpr7 and may thus require a conformational state of eEF2 which is maintained by specific (co-)chaperones.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-10DOI: 10.1002/1873-3468.15098
Ryuka Iizuka, Takahiro Ogawa, Rikako Tsukida, Keiichi Noguchi, David Hibbett, Yoko Katayama, Makoto Yoshida
{"title":"Characterization of fungal carbonyl sulfide hydrolase belonging to clade D β-carbonic anhydrase.","authors":"Ryuka Iizuka, Takahiro Ogawa, Rikako Tsukida, Keiichi Noguchi, David Hibbett, Yoko Katayama, Makoto Yoshida","doi":"10.1002/1873-3468.15098","DOIUrl":"https://doi.org/10.1002/1873-3468.15098","url":null,"abstract":"<p><p>Carbonyl sulfide hydrolase (COSase) is a unique enzyme that exhibits high activity towards carbonyl sulfide (COS) but low carbonic anhydrase (CA) activity, despite belonging to the CA family. COSase was initially identified in a sulfur-oxidizing bacterium and later discovered in the ascomycete Trichoderma harzianum strain THIF08. The COSase from T. harzianum has been recognized as a key enzyme in the assimilation of gaseous COS, and homologous genes are widely present not only in Ascomycota but also in Basidiomycota. Here, we characterized the COSases from the basidiomycete Gloeophyllum trabeum NBRC 6430 and T. harzianum to obtain detailed characteristics of fungal COSase. This study contributes to a better understanding of COS metabolism in fungi.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142962146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-07DOI: 10.1002/1873-3468.15078
Zhenan Liu, Joonho Yoon, Eunyoung Lee, Audrey N. Chang, R. Tyler Miller
{"title":"Calcium-sensing receptor- and ADAM10-mediated klotho shedding is regulated by tetraspanin 5","authors":"Zhenan Liu, Joonho Yoon, Eunyoung Lee, Audrey N. Chang, R. Tyler Miller","doi":"10.1002/1873-3468.15078","DOIUrl":"10.1002/1873-3468.15078","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <p>Soluble, circulating Klotho (sKlotho) is essential for normal health and renal function. sKlotho is shed from the renal distal convoluted tubule (DCT), its primary source, via enzymatic cleavage. However, the physiologic mechanisms that control sKlotho production, trafficking, and shedding are not fully defined. We previously found that the G protein-coupled calcium-sensing receptor (CaSR) co-localizes with membrane-bound αKlotho and the disintegrin/metalloprotease ADAM10 in the DCT and controls sKlotho in response to CaSR ligands and pHo by activating ADAM10. Here, we advance understanding of this process by showing that tetraspanin 5 (Tspan5), a scaffolding and chaperone protein, contributes to the cell surface expression and specificity of a protein complex that includes Tspan5, ADAM10, Klotho, and CaSR. These results support a model of multiprotein complexes that confer signaling specificity beyond CaSR on G protein-coupled processes.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <div>\u0000 <div>\u0000 \u0000 <h3>Impact statement</h3>\u0000 <p>Systemic circulating sKlotho is a determinant for normal physiology. Studies of knockout animals established its role as an anti-aging protein. The regulatory mechanisms for Klotho production and secretion are largely unknown. We report that Tspan 5 contributes to CaSR- and ADAM10-dependent Klotho shedding from the kidney, its primary source.</p>\u0000 </div>\u0000 </div>\u0000 </section>\u0000 </div>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":"599 6","pages":"866-875"},"PeriodicalIF":3.5,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1873-3468.15078","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142946818","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-05DOI: 10.1002/1873-3468.15077
Seraphine Kamayirese, Laura A. Hansen, Sándor Lovas
{"title":"Ligand recognition by 14-3-3 proteins requires negative charges but not necessarily phosphorylation","authors":"Seraphine Kamayirese, Laura A. Hansen, Sándor Lovas","doi":"10.1002/1873-3468.15077","DOIUrl":"10.1002/1873-3468.15077","url":null,"abstract":"<p>Protein–protein interactions involving 14-3-3 proteins regulate various cellular activities in normal and pathological conditions. These interactions have mostly been reported to be phosphorylation-dependent, but the 14-3-3 proteins also interact with unphosphorylated proteins. In this work, we investigated whether phosphorylation is required, or, alternatively, whether negative charges are sufficient for 14-3-3ε binding. We substituted the pThr residue of pT(502–510) peptide by residues with a varying number of negative charges and investigated the binding of the peptides to 14-3-3ε using MD simulations and biophysical methods. We demonstrated that at least one negative charge is required for the peptides to bind 14-3-3ε, although phosphorylation is not necessary, and that two negative charges are preferable for high affinity binding. This discovery opens up new approaches for designing peptide-based 14-3-3 protein inhibitors.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":"599 6","pages":"838-847"},"PeriodicalIF":3.5,"publicationDate":"2025-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1873-3468.15077","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142931102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-05DOI: 10.1002/1873-3468.15097
Thao Nghi Hoang, Meritxell Wu-Lu, Alberto Collauto, Peter-Leon Hagedoorn, Madalina Alexandru, Maike Henschel, Shahram Kordasti, Maria Andrea Mroginski, Maxie M Roessler, Kourosh H Ebrahimi
{"title":"The [2Fe-2S] cluster of mitochondrial outer membrane protein mitoNEET has an O<sub>2</sub>-regulated nitric oxide access tunnel.","authors":"Thao Nghi Hoang, Meritxell Wu-Lu, Alberto Collauto, Peter-Leon Hagedoorn, Madalina Alexandru, Maike Henschel, Shahram Kordasti, Maria Andrea Mroginski, Maxie M Roessler, Kourosh H Ebrahimi","doi":"10.1002/1873-3468.15097","DOIUrl":"https://doi.org/10.1002/1873-3468.15097","url":null,"abstract":"<p><p>The mitochondrial outer membrane iron-sulphur ([Fe-S]) protein mitoNEET has been extensively studied as a target of the anti-inflammatory and type-2 diabetes drug pioglitazone and as a protein affecting mitochondrial respiratory rate. Despite these extensive past studies, its molecular function has yet to be discovered. Here, we applied an interdisciplinary approach and discovered an explicit nitric oxide (NO) access site to the mitoNEET [2Fe-2S] cluster. We found that O<sub>2</sub> and pioglitazone block NO access to the cluster, suggesting a molecular function for the mitoNEET [2Fe-2S] cluster in mitochondrial signal transduction. Our discovery hints at a new pathway via which mitochondria can sense hypoxia through O<sub>2</sub> protection of the mitoNEET [2Fe-2S] cluster, a new paradigm in understanding the importance of [Fe-S] clusters for gasotransmitter signal transduction in eukaryotes.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142931103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-02DOI: 10.1002/1873-3468.15096
Annabella Nouel Barreto, Luis G. Cuello, Maria E. Zoghbi
{"title":"ABC transporter activity is affected by the size of lipid nanodiscs","authors":"Annabella Nouel Barreto, Luis G. Cuello, Maria E. Zoghbi","doi":"10.1002/1873-3468.15096","DOIUrl":"10.1002/1873-3468.15096","url":null,"abstract":"<p>Lipid nanodiscs have become a widely used approach for studying membrane proteins thanks to several advantages they offer. They have been especially useful for studying ABC transporters, despite the growing concern about the possible restriction of the conformational changes of the transporters due to the small size of the discs. Here, we performed a systematic study to determine the effect of the nanodisc size on the ATPase activity of model ABC transporters from human, plant, and bacteria. Our data confirm that the activity of the transporters and their response to regulatory molecules is affected by the nanodisc size. Our findings suggest the use of larger membrane scaffold proteins (MSPs), such as MSP2N2 nanodiscs, to minimize alterations caused by the commonly used small MSP1D1.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":"599 4","pages":"502-511"},"PeriodicalIF":3.5,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1873-3468.15096","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Specificity and sensitivity of ALT-associated markers in cancer cells.","authors":"Ion Udroiu, Jessica Marinaccio, Romina Stella Goffi, Emanuela Micheli, Antonella Sgura","doi":"10.1002/1873-3468.15087","DOIUrl":"https://doi.org/10.1002/1873-3468.15087","url":null,"abstract":"<p><p>Some tumors employ a mechanism called alternative lengthening of telomeres (ALT) to counteract telomere shortening-induced replicative senescence. Several hallmarks are used to identify cell lines and tumors as ALT-positive. Here, we analyzed a panel of ALT-positive and -negative cancer cell lines to investigate the specificity and sensibility of ALT-associated markers. We found that all the markers showed high sensitivity, indicating that cells not showing ALT markers are not ALT cells. Conversely, specificity varied significantly, i.e., many markers yield false positives. Detection of false positives may have influenced previous estimations of ALT incidence among tumors. Moreover, claims on the 'coexistence' of ALT and telomerase perhaps should be reconsidered. The findings prompt further study into the nature of these markers and their roles as either part of the ALT machinery or as by-products.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142913978","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-01DOI: 10.1002/1873-3468.15083
Lilian Koppensteiner, Layla Mathieson, Liam Neilson, Richard A. O'Connor, Ahsan R. Akram
{"title":"IFNγ and TNFα drive an inflammatory secretion profile in cancer-associated fibroblasts from human non-small cell lung cancer","authors":"Lilian Koppensteiner, Layla Mathieson, Liam Neilson, Richard A. O'Connor, Ahsan R. Akram","doi":"10.1002/1873-3468.15083","DOIUrl":"10.1002/1873-3468.15083","url":null,"abstract":"<p>Cancer-associated fibroblasts (CAFs) are the dominant nonmalignant component of the tumour microenvironment (TME). CAFs demonstrate a high level of inter- and intra-tumour heterogeneity in solid tumours, though the drivers of CAF subpopulations are not fully understood. Here, we demonstrate that non-small cell lung cancer (NSCLC) patient-derived CAFs upregulate the secretion of inflammatory cytokines (IL6, LIF, IL33, GM-CSF, IL1ra) and chemokines (CCL2, CCL3, CCL4, CCL20, CXCL8, CXCL9, CXCL10, CXCL11) in response to <i>in vitro</i> co-culture with anti-CD3/anti-CD28-stimulated peripheral blood mononuclear cells (PBMCs) via IFNγ and TNFα. Furthermore, T-cell-derived IFNγ inhibits CXCL12 secretion by CAFs <i>in vitro</i>. Our results highlight the ability of T-cell effector cytokines to modulate the CAF secretome in NSCLC.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":"599 5","pages":"713-723"},"PeriodicalIF":3.5,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1873-3468.15083","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142913918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2025-01-01DOI: 10.1002/1873-3468.15094
Peng Liu, Sitian He, Anouk Mentink, Pieter Hart, Yongjun Wu, Leon W. M. M. Terstappen, Pascal Jonkheijm, Michiel Stevens
{"title":"Silica-coated magnetic nanobeads in a flow enrichment target capture Halbach (FETCH) magnetic separation system for circulating tumor cell enrichment","authors":"Peng Liu, Sitian He, Anouk Mentink, Pieter Hart, Yongjun Wu, Leon W. M. M. Terstappen, Pascal Jonkheijm, Michiel Stevens","doi":"10.1002/1873-3468.15094","DOIUrl":"10.1002/1873-3468.15094","url":null,"abstract":"<p>Detecting circulating tumor cells (CTCs) is challenging due to their low presence and heterogeneity. Traditional methods using EpCAM-based separation struggle with CTCs that have undergone epithelial-mesenchymal transition, as this results in lower EpCAM expression. This study presents the use of silica-coated magnetic nanobeads functionalized with streptavidin for CTC capture. Using the FETCH magnetic separation system, we validated the capture efficiency of our beads on tumor cells with varying EpCAM expression. Our beads showed superior capture rates for LNCaP (97%), PC3-9 (91%), PC3 (23%), A549 (22%), and T24 (8%) cells compared to commercial MojoSort™ beads. Despite slightly higher nonspecific binding than CellSearch, our beads demonstrated improved sensitivity for EpCAMlow cells, suggesting they have promise for enhanced CTC capture.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":"599 5","pages":"724-738"},"PeriodicalIF":3.5,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1873-3468.15094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142913964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2024-12-31DOI: 10.1002/1873-3468.15089
Olga Zimmermannova, Martin Kubeš, Tereza Przeczková, Gal Masrati
{"title":"Residues R177 and S178 of the human Na+/H+ antiporter NHA2 are involved in its inhibition by the flavonoid phloretin","authors":"Olga Zimmermannova, Martin Kubeš, Tereza Przeczková, Gal Masrati","doi":"10.1002/1873-3468.15089","DOIUrl":"10.1002/1873-3468.15089","url":null,"abstract":"<p>The <i>Homo sapiens</i> Na<sup>+</sup>/H<sup>+</sup> antiporter NHA2 (<i>SLC9B2</i>) transports Na<sup>+</sup> or Li<sup>+</sup> in exchange for protons across cell membranes, and its dysfunction results in various pathologies. The activity of <i>Hs</i>NHA2 is specifically inhibited by the flavonoid phloretin. Using bioinformatic modeling, we predicted two amino acids (R177 and S178) as being important for the binding of phloretin to the <i>Hs</i>NHA2 molecule. Functional expression of <i>Hs</i>NHA2 in <i>Saccharomyces cerevisiae</i> and its site-directed mutagenesis revealed that while the R177T mutation resulted in an antiporter that was less sensitive to phloretin, the S178T mutation enhanced the inhibitory effect of phloretin on <i>Hs</i>NHA2. Our data corroborate the transport properties of <i>Hs</i>NHA2 and its interactions with an inhibitor and can be helpful for the development of new therapeutics targeting this antiporter and its pleiotropic physiological functions.</p>","PeriodicalId":12142,"journal":{"name":"FEBS Letters","volume":"599 6","pages":"901-911"},"PeriodicalIF":3.5,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1873-3468.15089","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142906833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}