Biochemical Journal最新文献_第5页

Inhibition of RIPK1 or RIPK3 kinase activity post ischemia-reperfusion reduces the development of chronic kidney injury. 缺血再灌注后抑制 RIPK1 或 RIPK3 激酶活性可减少慢性肾损伤的发生。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2025-01-22 DOI: 10.1042/BCJ20240569

Aspasia Pefanis, Anjan K Bongoni, Jennifer L McRae, Evelyn J Salvaris, Nella Fisicaro, James M Murphy, Francesco L Ierino, Peter J Cowan

{"title":"Inhibition of RIPK1 or RIPK3 kinase activity post ischemia-reperfusion reduces the development of chronic kidney injury.","authors":"Aspasia Pefanis, Anjan K Bongoni, Jennifer L McRae, Evelyn J Salvaris, Nella Fisicaro, James M Murphy, Francesco L Ierino, Peter J Cowan","doi":"10.1042/BCJ20240569","DOIUrl":"10.1042/BCJ20240569","url":null,"abstract":"Ischemia-reperfusion injury (IRI) occurs when the blood supply to an organ is temporarily reduced and then restored. Kidney IRI is a form of acute kidney injury (AKI), which often progresses to kidney fibrosis. Necroptosis is a regulated necrosis pathway that has been implicated in kidney IRI. Necroptotic cell death involves the recruitment of the RIPK1 and RIPK3 kinases and the activation of the terminal effector, the mixed lineage kinase domain-like (MLKL) pseudokinase. Phosphorylated MLKL causes cell death by plasma membrane rupture, driving 'necroinflammation'. Owing to their apical role in the pathway, RIPK1 and RIPK3 have been implicated in the development of kidney fibrosis. Here, we used a mouse model of unilateral kidney IRI to assess whether the inhibition of RIPK1 or RIPK3 kinase activity reduces AKI and the progression to kidney fibrosis. Mice treated with the RIPK1 inhibitor Nec-1s, either before or after IR, showed reduced kidney injury at 24 hr compared with controls, whereas no protection was offered by the RIPK3 inhibitor GSK´872. In contrast, treatment with either inhibitor from days 3 to 9 post-IR reduced the degree of kidney fibrosis at day 28. These findings further support the role of necroptosis in IRI and provide important validation for the contribution of both RIPK1 and RIPK3 catalytic activities in the progression of kidney fibrosis. Targeting the necroptosis pathway could be a promising therapeutic strategy to mitigate kidney disease following IR.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"73-86"},"PeriodicalIF":4.4,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142862779","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

In silico, in vitro and in vivo characterisation of thiamin binding proteins from plant seeds. 从植物种子中提取的硫胺素结合蛋白在体内和体外的特性。

IF 4.1 3区生物学

Biochemical Journal Pub Date : 2025-01-20 DOI: 10.1042/bcj20240429

Maria Faustino,Simon Strobbe,Raul Sanchez-Muñoz,Da Cao,Ratnesh C Mishra,Tiago Lourenço,Margarida Oliveira,Dominique Van Der Straeten

{"title":"In silico, in vitro and in vivo characterisation of thiamin binding proteins from plant seeds.","authors":"Maria Faustino,Simon Strobbe,Raul Sanchez-Muñoz,Da Cao,Ratnesh C Mishra,Tiago Lourenço,Margarida Oliveira,Dominique Van Der Straeten","doi":"10.1042/bcj20240429","DOIUrl":"https://doi.org/10.1042/bcj20240429","url":null,"abstract":"Thiamin, an essential micronutrient, is a cofactor for enzymes involved in the central carbon metabolism and amino acids pathways. Despite efforts to enhance thiamin content in rice by incorporating thiamin biosynthetic genes, increasing thiamin content in endosperm remains challenging, possibly due to a lack of thiamin stability and/or a local sink. The introduction of storage proteins has been successful in biofortification strategies and similar efforts targeting thiamin led to a 3-4-fold increase in white rice. However, only one thiamin-binding protein (TBP) sequence has been described in plants, more specifically from sesame seeds. Therefore, we aimed to identify and characterize TBPs, as well as to evaluate the effect of their expression on thiamin concentration, using an approach integrating in silico, in vitro and in vivo methods. We identified putative TBPs from Oryza sativa (rice), Fagopyrum esculentum (buckwheat) and Zea mays (maize) and pinpointed the thiamin-binding pockets through molecular docking. FeTBP and OsTBP contained one pocket with binding affinities similar to E. coli TBP, a well-characterized thiamin-binding protein, supporting their function. In vivo expression studies of TBPs in tobacco leaves and rice callus resulted in increased thiamin levels, with FeTBP and OsTBP showing the most pronounced effects. Additionally, thermal shift assays confirmed the thiamin binding capabilities of FeTBP and OsTBP, as observed by the significant increases in melting temperatures upon thiamin binding, indicating protein stabilization. These findings offer new insights into diversity and function of plant TBPs and highlight the potential of FeTBP and OsTBP to modulate thiamin levels in crop plants.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"24 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142991742","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

Specialized killing across the domains of life by the type VI secretion systems of Pseudomonas aeruginosa. 通过铜绿假单胞菌的VI型分泌系统在生命的各个领域进行专门杀伤。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2025-01-08 DOI: 10.1042/BCJ20230240

Jake Colautti, Steven D Kelly, John C Whitney

{"title":"Specialized killing across the domains of life by the type VI secretion systems of Pseudomonas aeruginosa.","authors":"Jake Colautti, Steven D Kelly, John C Whitney","doi":"10.1042/BCJ20230240","DOIUrl":"https://doi.org/10.1042/BCJ20230240","url":null,"abstract":"Type VI secretion systems (T6SSs) are widespread bacterial protein secretion machines that inject toxic effector proteins into nearby cells, thus facilitating both bacterial competition and virulence. Pseudomonas aeruginosa encodes three evolutionarily distinct T6SSs that each export a unique repertoire of effectors. Owing to its genetic tractability, P. aeruginosa has served as a model organism for molecular studies of the T6SS. However, P. aeruginosa is also an opportunistic pathogen and ubiquitous environmental organism that thrives in a wide range of habitats. Consequently, studies of its T6SSs have provided insight into the role these systems play in the diverse lifestyles of this species. In this review, we discuss recent advances in understanding the regulation and toxin repertoire of each of the three P. aeruginosa T6SSs. We argue that these T6SSs serve distinct physiological functions; whereas one system is a dedicated defensive weapon for interbacterial antagonism, the other two T6SSs appear to function primarily during infection. We find support for this model in examining the signalling pathways that control the expression of each T6SS and co-ordinate the activity of these systems with other P. aeruginosa behaviours. Furthermore, we discuss the effector repertoires of each T6SS and connect the mechanisms by which these effectors kill target cells to the ecological conditions under which their respective systems are activated. Understanding the T6SSs of P. aeruginosa in the context of this organism's diverse lifestyles will provide insight into the physiological roles these secretion systems play in this remarkably adaptable bacterium.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"482 1","pages":"1-15"},"PeriodicalIF":4.4,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142943579","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 MYO1B and MYO5B motor proteins and the sorting nexin SNX27 regulate apical targeting of membrane mucin MUC17 in enterocytes. MYO1B和MYO5B运动蛋白以及分选连接蛋白SNX27调控肠细胞中膜黏液蛋白MUC17的顶端靶向。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2025-01-08 DOI: 10.1042/BCJ20240204

Sofia Jäverfelt, Gustaf Hellsén, Izumi Kaji, James R Goldenring, Thaher Pelaseyed

{"title":"The MYO1B and MYO5B motor proteins and the sorting nexin SNX27 regulate apical targeting of membrane mucin MUC17 in enterocytes.","authors":"Sofia Jäverfelt, Gustaf Hellsén, Izumi Kaji, James R Goldenring, Thaher Pelaseyed","doi":"10.1042/BCJ20240204","DOIUrl":"10.1042/BCJ20240204","url":null,"abstract":"A dense glycocalyx, composed of the megaDalton-sized membrane mucin MUC17, coats the microvilli in the apical brush border of transporting intestinal epithelial cells, called enterocytes. The formation of the MUC17-based glycocalyx in the mouse small intestine occurs at the critical suckling-weaning transition. The glycocalyx extends 1 µm into the intestinal lumen and prevents the gut bacteria from directly attaching to the enterocytes. To date, the mechanism behind the positioning of MUC17 to the brush border is not known. Here, we show that the actin-based motor proteins MYO1B and MYO5B, and the sorting nexin SNX27, regulate apical targeting of MUC17 in enterocytes. We demonstrate that MUC17 turnover at the brush border is slow and controlled by MYO1B and SNX27. Furthermore, we report that MYO1B regulates MUC17 protein levels in enterocytes, whereas MYO5B specifically governs MUC17 levels at the brush border. Together, our results extend our understanding of the apical targeting of membrane mucins and provide mechanistic insights into how defective positioning of MUC17 renders enterocytes sensitive to bacterial challenges.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1-23"},"PeriodicalIF":4.4,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805927","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

Loss of peroxiredoxin 6 alters lipid composition and distribution resulting in increased sensitivity to ferroptosis. 过氧化物歧化酶 6 (PRDX6) 的缺失会改变脂质的组成和分布，从而增加对铁中毒的敏感性。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2024-12-23 DOI: 10.1042/BCJ20240445

Daniel J Lagal, Ángel Ortiz-Alcántara, José R Pedrajas, Brian McDonagh, J Antonio Bárcena, Raquel Requejo-Aguilar, C Alicia Padilla

{"title":"Loss of peroxiredoxin 6 alters lipid composition and distribution resulting in increased sensitivity to ferroptosis.","authors":"Daniel J Lagal, Ángel Ortiz-Alcántara, José R Pedrajas, Brian McDonagh, J Antonio Bárcena, Raquel Requejo-Aguilar, C Alicia Padilla","doi":"10.1042/BCJ20240445","DOIUrl":"10.1042/BCJ20240445","url":null,"abstract":"Peroxiredoxin 6 (PRDX6) is a multifunctional enzyme involved in phospholipid peroxide repair and metabolism. In this study we investigated the global lipid composition of a human hepatocarcinoma cell line SNU475 lacking PRDX6 and lipid related cellular processes. There was a general decrease in multiple lipids species upon loss of PRDX6, in particular sphingomyelins and acylcarnitines, consistent with previously observed alterations in cell signaling pathways and mitochondrial dysfunction. Deprivation of docosahexaenoic acid and related species was also evident. However, a few striking exceptions are worth highlighting: (1) Three specific arachidonic acid (AA) containing phophatidylcholines (PC) increased significantly. The increase of sn1-stearic/sn2-PUFA containing PC and sn2-AA containing plasmenyls are indicative of a preference of PRDX6 iPLA2 activity for these AA storage glycerophospholipids. (2) Several polyunsaturated fatty acids (PUFA) and PUFA containing triacylglycerols accumulated together with increased formation of lipid droplets, an indication of altered FA flux and PUFA sequestration in PRDX6 knockout cells. Loss of PRDX6 resulted in increased sensitivity to erastin-induced ferroptosis, independent of selenium and GPX4, as a consequence of increased levels of lipid hydroperoxides, that reverted to normal levels upon rescue with PRDX6. The results presented demonstrate that all three enzymatic activities of PRDX6 contribute to the role of this multifunctional enzyme in diverse cellular processes, including membrane phospholipid remodeling and glycerophospholipid functional diversity, resulting in altered lipid peroxides and modulation of AA disposition and traffic. These contributions highlight the complexity of the changes that loss of PRDX6 exerts on cell functionality.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1997-2015"},"PeriodicalIF":4.4,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11668489/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142725333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tweaking the redox properties of PpcA from Geobacter metallireducens with protein engineering. 用蛋白质工程技术调节金属还原杆菌PpcA的氧化还原特性。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2024-12-23 DOI: 10.1042/BCJ20240423

Pilar C Portela, Marta A Silva, Alexandre Almeida, Gonçalo F Damas, Carlos A Salgueiro

{"title":"Tweaking the redox properties of PpcA from Geobacter metallireducens with protein engineering.","authors":"Pilar C Portela, Marta A Silva, Alexandre Almeida, Gonçalo F Damas, Carlos A Salgueiro","doi":"10.1042/BCJ20240423","DOIUrl":"10.1042/BCJ20240423","url":null,"abstract":"Geobacter's unique ability to perform extracellular electron transfer (EET) to electrodes in microbial fuel cells (MFCs) has sparked the implementation of sustainable production of electrical energy. However, the electrochemical performance of Geobacter's biofilms in MFCs remains challenging to implement industrially. Multiple approaches are being investigated to enhance MFC technologies. Protein engineering of multihaem cytochromes, key components of Geobacter's EET pathways, can, conceivably, be pursued to improve the EET chain. The periplasmic cytochrome PpcA bridges ET from the inner to the outer membrane and its deletion impairs this crucial step. The functional characterisation of PpcA homologues from Geobacter sulfurreducens (Gs) and Geobacter metallireducens (Gm) revealed a significantly different redox behaviour even though they only differ by thirteen amino acids. In a previous study, we found that the single replacement of a tryptophan residue by methionine (W45M) in PpcAGm shifted the reduction potential value 33% towards that of PpcAGs. In this work, we expanded our investigation to include other non-conserved residues by conducting five mutation rounds. We identified the most relevant residues controlling the redox properties of PpcAGm. With just four mutations (K19, G25, N26, W45) the reduction potential value of PpcAGm was shifted 71% toward that of PpcAGs. Additionally, in the quadruple mutant, it was possible to replicate the haem oxidation order and the functional mechanisms of PpcAGs, which differ from those in PpcAGm. Overall, the mutants exhibit diverse redox and functional mechanisms that could be explored as a library for the future design of minimal, synthetic, ET chains in Geobacter.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"2017-2036"},"PeriodicalIF":4.4,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765734","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 open to closed D-loop conformational switch determines length in filopodia-like actin bundles. 打开到闭合的d环构象开关决定了丝状伪足样肌动蛋白束的长度。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2024-12-23 DOI: 10.1042/BCJ20240367

Jonathan R Gadsby, Pantelis Savvas Ioannou, Richard Butler, Julia Mason, Alison J Smith, Ulrich Dobramysl, Stacey E Chin, Claire Dobson, Jennifer L Gallop

{"title":"The open to closed D-loop conformational switch determines length in filopodia-like actin bundles.","authors":"Jonathan R Gadsby, Pantelis Savvas Ioannou, Richard Butler, Julia Mason, Alison J Smith, Ulrich Dobramysl, Stacey E Chin, Claire Dobson, Jennifer L Gallop","doi":"10.1042/BCJ20240367","DOIUrl":"10.1042/BCJ20240367","url":null,"abstract":"Filopodia, microspikes and cytonemes are implicated in sensing the environment and in dissemination of morphogens, organelles and pathogens across tissues. Their major structural component is parallel bundles of actin filaments that assemble from the cell membrane. Whilst the length of filopodia is central to their function, it is not known how their lengths are determined by actin bundle dynamics. Here, we identified a set of monoclonal antibodies that lengthen filopodia-like structures formed in a cell-free reconstitution system, and used them to uncover a key molecular switch governing length regulation. Using immunolabelling, enzyme-linked immunosorbent assays, immunoprecipitation and immunoblock experiments, we identified four antibodies that lengthen actin bundles by selectively binding the open DNase 1-binding loop (D-loop) of actin filaments. The antibodies inhibit actin disassembly and their effects can be alleviated by providing additional actin or cofilin. This work indicates that maintaining an open state of the actin filament D-loop is a mechanism of generating long filopodia-like actin bundles.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1977-1995"},"PeriodicalIF":4.4,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11668490/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of circadian rhythms in mammalian systems. 哺乳动物生理节律的发展。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2024-12-23 DOI: 10.1042/BCJ20210060

Junghyun Lee, Sevde Goker, Sookkyung Lim, Christian I Hong

引用次数: 0

Gα12 and Gα13 proteins are required for transforming growth factor-β-induced myofibroblast differentiation. TGF-β诱导的肌成纤维细胞分化需要Ga12和Ga13蛋白。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2024-12-18 DOI: 10.1042/BCJ20240317

Eleanor B Reed, Albert Sitikov, Kun Woo D Shin, Robert B Hamanaka, Rengül Cetin-Atalay, Gökhan M Mutlu, Alexander A Mongin, Nickolai O Dulin

{"title":"Gα12 and Gα13 proteins are required for transforming growth factor-β-induced myofibroblast differentiation.","authors":"Eleanor B Reed, Albert Sitikov, Kun Woo D Shin, Robert B Hamanaka, Rengül Cetin-Atalay, Gökhan M Mutlu, Alexander A Mongin, Nickolai O Dulin","doi":"10.1042/BCJ20240317","DOIUrl":"10.1042/BCJ20240317","url":null,"abstract":"Myofibroblast differentiation, characterized by accumulation of cytoskeletal and extracellular matrix proteins by fibroblasts, is a key process in wound healing and pathogenesis of tissue fibrosis. Transforming growth factor-β (TGF-β) is the most powerful known driver of myofibroblast differentiation. TGF-β signals through transmembrane receptor serine/threonine kinases that phosphorylate Smad transcription factors (Smad2/3) leading to activation of transcription of target genes. Heterotrimeric G proteins mediate distinct signaling from seven-transmembrane G protein coupled receptors, which are not known to be linked to Smad activation. We tested whether G protein signaling plays any role in TGF-β-induced myofibroblast differentiation, using primary cultured human lung fibroblasts. Activation of Gαs by cholera toxin blocked TGF-β-induced myofibroblast differentiation without affecting Smad2/3 phosphorylation. Neither inhibition of Gαi by pertussis toxin nor siRNA-mediated combined knockdown of Gαq and Gα11 had a significant effect on TGF-β-induced myofibroblast differentiation. In contrast, combined knockdown of Gα12 and Gα13 significantly inhibited TGF-β-stimulated expression of myofibroblast marker proteins (collagen-1, fibronectin, smooth-muscle α-actin), with siGα12 being significantly more potent than siGα13. Mechanistically, combined knockdown of Gα12 and Gα13 resulted in substantially reduced phosphorylation of Smad2 and Smad3 in response to TGF-β, which was accompanied by a significant decrease in the expression of TGF-β receptors (TGFBR1, TGFBR2) and of Smad3. Thus, our study uncovers a novel role of Gα12/13 proteins in the control of TGF-β signaling and myofibroblast differentiation.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1937-1948"},"PeriodicalIF":4.4,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11668492/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

On the function of TRAP substrate-binding proteins: the isethionate-specific binding protein IseP. 关于 TRAP 底物结合蛋白的功能：异蛋氨酸特异性结合蛋白 IseP。

IF 4.4 3区生物学

Biochemical Journal Pub Date : 2024-12-18 DOI: 10.1042/BCJ20240540

Michael C Newton-Vesty, Michael J Currie, James S Davies, Santosh Panjikar, Ashish Sethi, Andrew E Whitten, Zachary D Tillett, David M Wood, Joshua D Wright, Michael J Love, Timothy M Allison, Sam A Jamieson, Peter D Mace, Rachel A North, Renwick C J Dobson

{"title":"On the function of TRAP substrate-binding proteins: the isethionate-specific binding protein IseP.","authors":"Michael C Newton-Vesty, Michael J Currie, James S Davies, Santosh Panjikar, Ashish Sethi, Andrew E Whitten, Zachary D Tillett, David M Wood, Joshua D Wright, Michael J Love, Timothy M Allison, Sam A Jamieson, Peter D Mace, Rachel A North, Renwick C J Dobson","doi":"10.1042/BCJ20240540","DOIUrl":"10.1042/BCJ20240540","url":null,"abstract":"Bacteria evolve mechanisms to compete for limited resources and survive in new niches. Here we study the mechanism of isethionate import from the sulfate-reducing bacterium Oleidesulfovibrio alaskensis. The catabolism of isethionate by Desulfovibrio species has been implicated in human disease, due to hydrogen sulfide production, and has potential for industrial applications. O. alaskensis employs a tripartite ATP-independent periplasmic (TRAP) transporter (OaIsePQM) to import isethionate, which relies on the substrate-binding protein (OaIseP) to scavenge isethionate and deliver it to the membrane transporter component (OaIseQM) for import into the cell. We determined the binding affinity of isethionate to OaIseP by isothermal titration calorimetry, KD = 0.95 µM (68% CI = 0.6-1.4 µM), which is weaker compared with other TRAP substrate-binding proteins. The X-ray crystal structures of OaIseP in the ligand-free and isethionate-bound forms were obtained and showed that in the presence of isethionate, OaIseP adopts a closed conformation whereby two domains of the protein fold over the substrate. We serendipitously discovered two crystal forms with sulfonate-containing buffers (HEPES and MES) bound in the isethionate-binding site. However, these do not evoke domain closure, presumably because of the larger ligand size. Together, our data elucidate the molecular details of how a TRAP substrate-binding protein binds a sulfonate-containing substrate, rather than a typical carboxylate-containing substrate. These results may inform future antibiotic development to target TRAP transporters and provide insights into protein engineering of TRAP transporter substrate-binding proteins.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1901-1920"},"PeriodicalIF":4.4,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11668362/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142667211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0