Journal of Biological Chemistry最新文献_第6页

Molecular insights into the structure and function of the Staphylococcus aureus fatty acid kinase. 金黄色葡萄球菌脂肪酸激酶结构和功能的分子见解。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107920

Megan J Myers, Zhen Xu, Benjamin J Ryan, Zachary R DeMars, Miranda J Ridder, David K Johnson, Christina N Krute, Tony S Flynn, Maithri M Kashipathy, Kevin P Battaile, Nicholas Schnicker, Scott Lovell, Bret D Freudenthal, Jeffrey L Bose

{"title":"Molecular insights into the structure and function of the Staphylococcus aureus fatty acid kinase.","authors":"Megan J Myers, Zhen Xu, Benjamin J Ryan, Zachary R DeMars, Miranda J Ridder, David K Johnson, Christina N Krute, Tony S Flynn, Maithri M Kashipathy, Kevin P Battaile, Nicholas Schnicker, Scott Lovell, Bret D Freudenthal, Jeffrey L Bose","doi":"10.1016/j.jbc.2024.107920","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107920","url":null,"abstract":"Gram-positive bacteria utilize a Fatty Acid Kinase (FAK) complex to harvest fatty acids from the environment. This complex consists of the fatty acid kinase, FakA, and an acyl carrier protein, FakB, and is known to impact virulence and disease outcomes. Despite some recent studies, there remains many outstanding questions as to the enzymatic mechanism and structure of FAK . To better address this gap in knowledge, we used a combination of modeling, biochemical, and cell-based approaches to build on prior proposed models and identify critical details of FAK activity. Using bio-layer interferometry, we demonstrated nanomolar affinity between FakA and FakB that also indicates that FakA is dimer when binding FakB. Additionally, targeted mutagenesis of the FakA Middle domain demonstrates it possesses a metal binding pocket that is critical for FakA dimer stability and FAK function in vitro and in vivo. Lastly, we solved structures of the apo and ligand-bound FakA kinase domain to capture the molecular changes in the protein following ATP binding and hydrolysis. Together, these data provide critical insight into the structure and function of the FAK complex which is essential for understanding its mechanism.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500927","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 nuclear transportation of CHRONO regulates the circadian rhythm. CHRONO 的核运输调节昼夜节律。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107917

Qin Zhou, Yunxia Su, Ruohan Wang, Zhiyuan Song, Honghua Ge, Ximing Qin

{"title":"The nuclear transportation of CHRONO regulates the circadian rhythm.","authors":"Qin Zhou, Yunxia Su, Ruohan Wang, Zhiyuan Song, Honghua Ge, Ximing Qin","doi":"10.1016/j.jbc.2024.107917","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107917","url":null,"abstract":"The pace of the endogenous circadian clock is important for organisms to maintain homeostasis. CHRONO has been shown to be a core component of the mammalian clock and has recently been implicated to function in several important physiological aspects. To function properly, CHRONO needs to enter the nucleus to repress transcription. We have previously shown that the N-terminus of CHRONO is required for its nuclear entry. However, how CHRONO enters the nucleus and regulates the circadian clock remains unknown. Here, we report that a novel non-classical nuclear localization signal (NLS) in the N-terminus of CHRONO is responsible for its nuclear entry. Multiple nuclear transporters are identified that facilitate the nuclear import of CHRONO. We show that the Arg63 is the critical amino acid of the NLS. Using prime editing technology, we precisely edit the Arg63 to Ala at the genomic loci and demonstrate that this mutation prolongs the circadian period, which is similar to knockdown of CHRONO. By using the CHRONO knockout and R63A mutant cells, we also investigated the changes in the cytoplasmic/nuclear distribution of BMAL1. We show that BMAL1 localizes more in the cytoplasm in the deficiency of CHRONO nuclear entry. These results provide a model for CHRONO nuclear entry using a network of importins involved in the regulation of the circadian period.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500960","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 crystal structure of Grindelia robusta 7,13-copalyl diphosphate synthase reveals active site features controlling catalytic specificity. Grindelia robusta 7,13-醛基二磷酸合酶的晶体结构揭示了控制催化特异性的活性位点特征。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107921

Anna E Cowie, Jose H Pereira, Andy DeGiovanni, Ryan P McAndrew, Malathy Palayam, Jedidiah O Peek, Andrew J Muchlinski, Yasuo Yoshikuni, Nitzan Shabek, Paul D Adams, Philipp Zerbe

{"title":"The crystal structure of Grindelia robusta 7,13-copalyl diphosphate synthase reveals active site features controlling catalytic specificity.","authors":"Anna E Cowie, Jose H Pereira, Andy DeGiovanni, Ryan P McAndrew, Malathy Palayam, Jedidiah O Peek, Andrew J Muchlinski, Yasuo Yoshikuni, Nitzan Shabek, Paul D Adams, Philipp Zerbe","doi":"10.1016/j.jbc.2024.107921","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107921","url":null,"abstract":"Diterpenoid natural products serve critical functions in plant development and ecological adaptation and many diterpenoids have economic value as bioproducts. The family of class II diterpene synthases catalyzes the committed reactions in diterpenoid biosynthesis, converting a common geranylgeranyl diphosphate precursor into different bicyclic prenyl diphosphate scaffolds. Enzymatic rearrangement and modification of these precursors generates the diversity of bioactive diterpenoids. We report the crystal structure of Grindelia robusta 7,13-copalyl diphosphate synthase, GrTPS2, at 2.1 Å of resolution. GrTPS2 catalyzes the committed reaction in the biosynthesis of grindelic acid, which represents the signature metabolite in species of gumweed (Grindelia spp., Asteraceae). Grindelic acid has been explored as a potential source for drug leads and biofuel production. The GrTPS2 crystal structure adopts the conserved three-domain fold of class II diterpene synthases featuring a functional active site in the γβ-domain and a vestigial ɑ-domain. Substrate docking into the active site of the GrTPS2 apo protein structure predicted catalytic amino acids. Biochemical characterization of protein variants identified residues with impact on enzyme activity and catalytic specificity. Specifically, mutagenesis of Y457 provided mechanistic insight into the position-specific deprotonation of the intermediary carbocation to form the characteristic 7,13 double bond of 7,13-copalyl diphosphate.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500959","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

Identification of a Staphylococcal dipeptidase involved in the production of human body odour. 鉴定参与产生人体气味的葡萄球菌二肽酶。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107928

Reyme Herman, Bethan Kinniment-Williams, Michelle Rudden, Alexander Gordon James, Anthony J Wilkinson, Barry Murphy, Gavin H Thomas

{"title":"Identification of a Staphylococcal dipeptidase involved in the production of human body odour.","authors":"Reyme Herman, Bethan Kinniment-Williams, Michelle Rudden, Alexander Gordon James, Anthony J Wilkinson, Barry Murphy, Gavin H Thomas","doi":"10.1016/j.jbc.2024.107928","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107928","url":null,"abstract":"The production of human body odour is the result of the action of commensal skin bacteria, including Staphylococcus hominis, acting to biotransform odourless apocrine gland secretions into volatile chemicals like thioalcohols such as 3-methyl-3-sulphanylhexan-1-ol (3M3SH). As the secreted odour precursor Cys-Gly-3M3SH contains a dipeptide, yet the final enzyme in the biotransformation pathway only functions on Cys-3M3SH, we sought to identify the remaining step in this human-adapted biochemical pathway using a novel coupled enzyme assay. Purification of this activity from S. hominis extracts led to the identification of the M20A-family PepV peptidase (ShPepV) as the primary Cys-Gly-3M3SH dipeptidase. To establish whether this was a primary substrate for PepV, the recombinant protein was purified and demonstrated broad activity against diverse dipeptides. The binding site for Cys-Gly-3M3SH was predicted using modelling, which suggested mutations that might accommodate this ligand more favourably. Indeed, a D437A resulted in an almost 6-fold increase in the kcat/KM, while other introduced mutations reduced or abolished function. Together these data identify an enzyme capable of catalysing the missing step in an ancient human-specific biochemical transformation and suggest that the production of 3M3SH neither uses a dedicated transporter nor peptidase for its breakdown, with only the final cleavage step, catalysed by PatB C-S β-lyase, being a unique enzyme.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500926","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

How to target membrane proteins for degradation: Bringing GPCRs into the TPD fold. 如何靶向降解膜蛋白：将 GPCR 纳入 TPD 框架。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107926

Boguslawa Korona, Laura S Itzhaki

引用次数: 0

DNA damage-induced p53 downregulates expression of RAG1 through a negative feedback loop involving miR-34a and FOXP1. DNA 损伤诱导的 p53 通过涉及 miR-34a 和 FOXP1 的负反馈回路下调 RAG1 的表达。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107922

Katarina Ochodnicka-Mackovicova, Martine van Keimpema, Marcel Spaargaren, Carel J M van Noesel, Jeroen E J Guikema

{"title":"DNA damage-induced p53 downregulates expression of RAG1 through a negative feedback loop involving miR-34a and FOXP1.","authors":"Katarina Ochodnicka-Mackovicova, Martine van Keimpema, Marcel Spaargaren, Carel J M van Noesel, Jeroen E J Guikema","doi":"10.1016/j.jbc.2024.107922","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107922","url":null,"abstract":"During the maturation of pre-B cells, the recombination activating gene 1 and 2 (RAG1/2) endonuclease complex plays a crucial role in coordinating V(D)J recombination by introducing DNA breaks in immunoglobulin (Ig) loci. Dysregulation of RAG1/2 has been linked to the onset of B-cell malignancies, yet the mechanisms controlling RAG1/2 in pre-B cells exposed to excessive DNA damage are not fully understood. In this study, we show that DNA damage-induced activation of p53 initiates a negative-feedback loop which rapidly downregulates RAG1 levels. This feedback loop involves ataxia telangiectasia mutated (ATM) activation, subsequent stabilization of p53, and modulation of microRNA-34a (miR-34a) levels, which is one of the p53 targets. Notably, this loop incorporates transcription factor forkhead box P1 (FOXP1) as a downstream effector. The absence of p53 resulted in an increased proportion of IgM+ cells prompted to upregulate RAG1/2 and to undergo Ig light chain (Igl) recombination. Similar results were obtained in primary pre-B cells with depleted levels of miR-34a. We propose that in pre-B cells undergoing Ig gene recombination, the DNA breaks activate a p53/miR-34a/FOXP1-mediated negative-feedback loop that contributes to the rapid downregulation of RAG. This regulation limits the RAG-dependent DNA damage, thereby protecting the stability of the genome during V(D)J rearrangement in developing B cells.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500921","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

DNA polymerase ζ has robust reverse transcriptase activity relative to other cellular DNA polymerases. 与其他细胞 DNA 聚合酶相比，DNA 聚合酶 ζ 具有强大的逆转录酶活性。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107918

Ryan Mayle, William K Holloman, Michael E O'Donnell

{"title":"DNA polymerase ζ has robust reverse transcriptase activity relative to other cellular DNA polymerases.","authors":"Ryan Mayle, William K Holloman, Michael E O'Donnell","doi":"10.1016/j.jbc.2024.107918","DOIUrl":"10.1016/j.jbc.2024.107918","url":null,"abstract":"Cell biology and genetic studies have demonstrated that DNA double strand break (DSB) repair can be performed using an RNA transcript that spans the site of the DNA break as a template for repair. This type of DSB repair requires a reverse transcriptase to convert an RNA sequence into DNA to facilitate repair of the break, rather than copying from a DNA template as in canonical DSB repair. Translesion synthesis (TLS) DNA polymerases (Pol) are often more promiscuous than DNA Pols, raising the notion that reverse transcription could be performed by a TLS Pol. Indeed, several studies have demonstrated that human Pol η has reverse transcriptase activity, while others have suggested that the yeast TLS Pol ζ is involved. Here, we purify all seven known nuclear DNA Pols of Saccharomyces cerevisiae and compare their reverse transcriptase activities. The comparison shows that Pol ζ far surpasses Pol η and all other DNA Pols in reverse transcriptase activity. We find that Pol ζ reverse transcriptase activity is not affected by RPA or RFC/PCNA and acts distributively to make DNA complementary to an RNA template strand. Consistent with prior S. cerevisiae studies performed in vivo, we propose that Pol ζ is the major DNA Pol that functions in the RNA templated DSB repair pathway.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500922","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

Hepcidin expression is associated with increased γ-secretase-mediated cleavage of neogenin in the liver. 肝素的表达与肝脏中γ-分泌酶介导的新肝素裂解增加有关。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-23 DOI: 10.1016/j.jbc.2024.107927

Caroline A Enns, Richard H Zhang, Shall Jue, An-Sheng Zhang

{"title":"Hepcidin expression is associated with increased γ-secretase-mediated cleavage of neogenin in the liver.","authors":"Caroline A Enns, Richard H Zhang, Shall Jue, An-Sheng Zhang","doi":"10.1016/j.jbc.2024.107927","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107927","url":null,"abstract":"Neogenin (NEO1) is a ubiquitously expressed transmembrane protein. It interacts with hemojuvelin (HJV). Both NEO1 and HJV play pivotal roles in iron homeostasis by inducing hepcidin expression in the liver. Our previous studies demonstrated that this process depends on Neo1-Hjv interaction and showed that the Hjv-mediated hepcidin expression is correlated with the accumulation of a truncated and membrane-associated form of Neo1. In this study, we tested whether hepcidin expression is induced by increased γ-secretase-mediated cleavage of Neo1 in the liver. We found that Neo1 underwent cleavage of its ectodomain and intracellular domains by α- and γ-secretases, respectively, in hepatoma cells. Our in vitro studies suggest that γ-secretase is responsible for cleavage and release of the cytoplasmic domain of Neo1 in the Hjv-Neo1 complex. This process was enhanced by inhibition of α-secretase proteolysis and by co-expression with the Neo1-binding partner, Alk3. Further in vivo studies indicated that Neo1 induction of hepcidin expression required γ-secretase cleavage. Interestingly, neither predicted form of γ-secretase-cleaved Neo1 was able to induce hepcidin when separately expressed in hepatocyte-specific Neo1 knockout mice. These results imply that the function of Neo1 requires a de novo γ-secretase proteolysis. Additional studies revealed that in addition to the Hjv-binding domains, the function of Neo1 also required its C-terminal intracellular domain and the N-terminal immunoglobulin-like domains that are involved in Neo1 binding to Alk3. Together, our data support the idea that Neo1 induction of hepcidin is initiated as a full-length form and requires a de novo γ-secretase cleavage of Neo1's cytoplasmic domain.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500924","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

Identification of the N-terminal residues responsible for the differential microdomain localization of CYP1A1 and CYP1A2. 鉴定导致 CYP1A1 和 CYP1A2 不同微域定位的 N 端残基。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-22 DOI: 10.1016/j.jbc.2024.107891

Robert M Fuchs,James R Reed,J Patrick Connick,Markéta Paloncýová,Martin Šrejber,Petra Čechová,Michal Otyepka,Marilyn K Eyer,Wayne L Backes

{"title":"Identification of the N-terminal residues responsible for the differential microdomain localization of CYP1A1 and CYP1A2.","authors":"Robert M Fuchs,James R Reed,J Patrick Connick,Markéta Paloncýová,Martin Šrejber,Petra Čechová,Michal Otyepka,Marilyn K Eyer,Wayne L Backes","doi":"10.1016/j.jbc.2024.107891","DOIUrl":"https://doi.org/10.1016/j.jbc.2024.107891","url":null,"abstract":"The endoplasmic reticulum (ER) is organized into ordered regions enriched in cholesterol and sphingomyelin, and disordered microdomains characterized by more fluidity. Rabbit CYP1A1 and CYP1A2 localize into disordered and ordered microdomains, respectively. Previously, a CYP1A2 chimera containing the first 109 amino acids of CYP1A1 showed altered microdomain localization. The goal of this study was to identify specific residues responsible for CYP1A microdomain localization. Thus, CYP1A2 chimeras containing substitutions from homologous regions of CYP1A1 were expressed in HEK 293T/17 cells, and the localization was examined after solubilization with Brij 98. A CYP1A2 mutant with the three amino acids from CYP1A1 (VAG) at positions 27-29 of CYP1A2 was generated that showed a distribution pattern similar to those of CYP1A1/1A2 chimeras containing both the first 109 amino acids and the first 31 amino acids of CYP1A1 followed by remaining amino acids of CYP1A2. Similarly, the reciprocal substitution of three amino acids from CYP1A2 (AVR) into CYP1A1 resulted in a partial redistribution of the chimera into ordered microdomains. Molecular dynamic simulations indicate that the positive charges of the CYP1A1 and CYP1A2 linker regions between the N-termini and catalytic domains resulted in different depths of immersion of the N-termini in the membrane. The overlap of the distribution of positively charged residues in CYP1A2 (AVR) and negatively charged phospholipids was higher in the ordered than disordered microdomain. These findings identify three residues in the CYP1A N-terminus as a novel microdomain-targeting motif of the P450s and provide a mechanistic explanation for the differential microdomain localization of CYP1A.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142490377","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

Azotobacter vinelandii scaffold protein NifU transfers iron to NifQ as part of the iron-molybdenum cofactor biosynthesis pathway for nitrogenase. 作为氮酶铁钼辅助因子生物合成途径的一部分，Azotobacter vinelandii 支架蛋白 NifU 将铁转移到 NifQ 上。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2024-10-22 DOI: 10.1016/j.jbc.2024.107900

Emma Barahona, Juan Andrés Collantes-García, Elena Rosa-Núñez, Jin Xiong, Xi Jiang, Emilio Jiménez-Vicente, Carlos Echávarri-Erasun, Yisong Guo, Luis M Rubio, Manuel González-Guerrero

{"title":"Azotobacter vinelandii scaffold protein NifU transfers iron to NifQ as part of the iron-molybdenum cofactor biosynthesis pathway for nitrogenase.","authors":"Emma Barahona, Juan Andrés Collantes-García, Elena Rosa-Núñez, Jin Xiong, Xi Jiang, Emilio Jiménez-Vicente, Carlos Echávarri-Erasun, Yisong Guo, Luis M Rubio, Manuel González-Guerrero","doi":"10.1016/j.jbc.2024.107900","DOIUrl":"10.1016/j.jbc.2024.107900","url":null,"abstract":"The Azotobacter vinelandii molybdenum nitrogenase obtains molybdenum from NifQ, a monomeric iron-sulfur molybdoprotein. This protein requires an existing [Fe-S] cluster to form a [Mo-Fe3-S4] group, which acts as a specific molybdenum donor during nitrogenase FeMo-co biosynthesis. Here, we show biochemical evidence supporting the role of NifU as the [Fe-S] cluster donor. Protein-protein interaction studies involving apo-NifQ and as-isolated NifU demonstrated their interaction, which was only effective when NifQ lacked its [Fe-S] cluster. Incubation of apo-NifQ with [Fe4-S4]-loaded NifU increased the iron content of the former, contingent on both proteins being able to interact with one another. As a result of this interaction, a [Fe4-S4] cluster was transferred from NifU to NifQ. In A. vinelandii, NifQ was preferentially metalated by NifU rather than by the [Fe-S] cluster scaffold protein IscU. These results indicate the necessity of co-expressing NifU and NifQ to efficiently provide molybdenum for FeMo-co biosynthesis when engineering nitrogenase in plants.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142500917","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