Jiannan Wang, Xiaogang Niu, Changwen Jin, Yunfei Hu
{"title":"<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of the third intracellular loop of the muscarinic acetylcholine receptor M1.","authors":"Jiannan Wang, Xiaogang Niu, Changwen Jin, Yunfei Hu","doi":"10.1007/s12104-025-10230-9","DOIUrl":"https://doi.org/10.1007/s12104-025-10230-9","url":null,"abstract":"<p><p>G protein-coupled receptors (GPCRs) are highly dynamic seven-transmembrane (7TM) proteins that respond to various extracellular stimuli and elicit diverse intracellular signaling cascades. The third intracellular loops (ICL3s) of the GPCRs are intrinsically disordered and play important roles in signaling. The muscarinic acetylcholine receptors (mAChRs) harbor extremely long ICL3s, which comprise over a hundred amino acid residues and contain multiple phosphorylation sites. Due to their intrinsic flexibility, ICL3s are commonly absent or unobservable in cryo-EM or X-ray structures, and there has been a lack of structural and dynamics study of these regions. Herein, we report the <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N chemical shift assignments of the M1 muscarinic receptor ICL3, which provides a basis for further NMR studies of its conformational dynamics, post-translational modifications and interactions.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770846","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}
Christina Muhs, Lena Kemper, Christian Richter, Francesca Lavore, Markus Weingarth, Anna Wacker, Harald Schwalbe
{"title":"NMR characterisation of the antibiotic resistance-mediating 32mer RNA from the 23S ribosomal RNA.","authors":"Christina Muhs, Lena Kemper, Christian Richter, Francesca Lavore, Markus Weingarth, Anna Wacker, Harald Schwalbe","doi":"10.1007/s12104-025-10229-2","DOIUrl":"https://doi.org/10.1007/s12104-025-10229-2","url":null,"abstract":"<p><p>The increasing prevalence of antibiotic resistance represents a significant public health concern, underscoring the urgent need for the development of novel therapeutic strategies. The antibiotic effects of macrolides, the second most widely used class of antibiotics, are counteracted by Erm proteins through the methylation of adenosine 2058 of the 23S ribosomal RNA (rRNA) (~ 2900 nucleotides), yielding either monomethylated or dimethylated A2058. This methylation is the molecular basis for preventing macrolides from binding and leads to the development of resistance of bacteria including Staphylococcus, Streptococcus and Enterococcus. While the function of Erm proteins have been thoroughly investigated, the role of the ribosomal RNA in acquiring antibiotic resistance is frequently underestimated, given that the ribosomal RNA is the actual target for methylation. Here, we present the comprehensive <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N NMR resonance assignment for the part of the 23S rRNA that serves as the Erm substrate in antimicrobial resistance. Furthermore, we compare the chemical shift signature of the unmethylated to the monomethylated and dimethylated RNA construct and show that changes in the RNA upon methylation are locally restricted. The resonance assignments provide a starting point for investigating and targeting the molecular mechanism of the resistance-conferring Erm proteins.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770870","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}
{"title":"Backbone resonance assignments of the CPEB3 [101-200] and CPEB3 [294-410].","authors":"Harunobu Saito, Yujin Lee, Tomoharu Ueno, Naotaka Sekiyama, Masatomo So, Ayako Furukawa, Kenji Sugase","doi":"10.1007/s12104-025-10226-5","DOIUrl":"https://doi.org/10.1007/s12104-025-10226-5","url":null,"abstract":"<p><p>Cytoplasmic polyadenylation element-binding protein 3 (CPEB3) is an RNA-binding protein that plays a pivotal role in the formation of long-term memory. The N-terminal region (residues 1-459) of CPEB3 is a highly aggregative intrinsically disordered region (IDR) that regulates the translation of specific targets, such as AMPA subunits, through mechanisms including liquid-liquid phase separation (LLPS) and the formation of fibrous aggregates. Despite its significance, the underlying mechanisms remain poorly understood. In this study, we present the backbone resonance assignments of residues 101-200 and 294-410 segments of the CPEB3 IDR. In agreement with sequence-based predictions, CPEB3 [101-200] was found to be disordered, whereas two partial α-helices were identified within CPEB3 [294-410].</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143741959","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}
Antoine Baudin, Hoang H Dinh, Xiaoping Xu, David S Libich
{"title":"The <sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C backbone resonance assignments of the N-terminal (1-149) domain of Serpine mRNA Binding Protein 1 (SERBP1).","authors":"Antoine Baudin, Hoang H Dinh, Xiaoping Xu, David S Libich","doi":"10.1007/s12104-025-10225-6","DOIUrl":"https://doi.org/10.1007/s12104-025-10225-6","url":null,"abstract":"<p><p>Serpine mRNA-Binding Protein 1 (SERBP1) is an RNA-binding protein implicated in diverse cellular functions, including translational regulation, tumor progression, and stress response. It interacts with ribosomal subunits, RNA, and proteins involved in stress granules, contributing to processes such as phase separation and epigenetic regulation. Recent studies have shown SERBP1's role in glioblastoma progression and its involvement in ribosomal regulation. Structurally, SERBP1 contains N- and C-terminal hyaluronan-binding domains, two RG/RGG motifs, and is predicted to be predominantly disordered. Here, we report the backbone resonance assignment and secondary structure propensities of SERBP1's N-terminal residues (1-149). Using NMR spectroscopy, we identified a stable α-helix (residues 28-40) and transient structural elements. These findings provide insight into the structural features of SERBP1 that may mediate its interactions with ribosomal subunits, RNA, and other binding partners, laying a foundation for future structural studies of its functional mechanisms.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735585","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}
Priti Chanda Behera, Sneha Paturi, Mandar V Deshmukh
{"title":"Chemical shift assignments of N-terminal dsRNA binding domains dsRBD1 and dsRBD2 of Arabidopsis thaliana DRB5.","authors":"Priti Chanda Behera, Sneha Paturi, Mandar V Deshmukh","doi":"10.1007/s12104-025-10224-7","DOIUrl":"https://doi.org/10.1007/s12104-025-10224-7","url":null,"abstract":"<p><p>Double-stranded RNA (dsRNA) binding proteins (dsRBPs) are among the key players that act along with other components involved in the RNA interference (RNAi) pathway for mediating gene silencing. Additionally, members of the dsRBP family of proteins play divergent roles in the broader array of biological processes. In Arabidopsis thaliana, dsRNA binding protein 5 (DRB5), along with DRB2 and DRB3, serves in recognition of viral RNA invasion and co-localizes with viral replication complexes. However, the functional role of DRB5 in such complexes is yet to be explored. DRB5 is a multidomain protein containing two tandem dsRNA binding domains (dsRBDs) at its N-terminus. Our current study presents the near-complete backbone and sidechain assignment of the dsRBD1 and dsRBD2 of DRB5 using solution NMR. The study will further contribute to determining the solution structure of dsRBDs and open new avenues to investigate the functional role of DRB5 in gene silencing pathways.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699391","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}
{"title":"Backbone resonance assignments of dopamine N-acetyltransferase in free and cofactor-bound states.","authors":"Chu-Ya Wu, Yi-Zong Lee, I-Chen Hu, Liang-Yuan Chiu, Wei-Cheng Ding, Jing Wang, Shih-Che Sue, Shin-Ichi Tate, Ping-Chiang Lyu","doi":"10.1007/s12104-025-10222-9","DOIUrl":"https://doi.org/10.1007/s12104-025-10222-9","url":null,"abstract":"<p><p>Dopamine N-acetyltransferase (Dat), belonging to the GCN5-related N-acetyltransferase (GNAT) superfamily, is an arylalkylamine N-acetyltransferase (AANAT) that is involved in insects neurotransmitter inactivation and the development of insect cuticle sclerotization. By using the cofactor acetyl coenzyme A (Ac-CoA) as an acetyl group donor, Dat produces acetyl-dopamine through the reaction with dopamine. Although AANATs share similar structural features with the GNAT family, they have low sequence identities among insect AANATs (~ 40%) and between insect AANATs and vertebrate AANATs (~ 12%). A common noticed feature in GNATs is the Ac-CoA-binding induced conformational change, and this is important for further selection and catalysis of its substrate. In AANATs, the conformational changes help the sequential binding mechanism. Here, we report the <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N backbone resonance assignments of the 24 kDa Dat from Drosophila melanogaster in the free and Ac-CoA-bound states, and the chemical shift differences revealed a significant conformational change in the α1 region of Dat. These assignments provide a foundation for further investigations of the catalysis and structural regulation of Dat in solution.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143397713","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}
{"title":"Backbone resonance assignment of the catalytic and ATP-binding domain of CpxA from Escherichia coli.","authors":"Jing Deng, Guofang Zeng, Wenqing Xia, Wei Tang, Zhaofei Chai, Yixiang Liu, Conggang Li, Liqun Huang, Ling Jiang","doi":"10.1007/s12104-025-10218-5","DOIUrl":"10.1007/s12104-025-10218-5","url":null,"abstract":"<p><p>CpxA is an extensively studied histidine kinase implicated in cellular stress responses. The highly conserved CA domain of CpxA (CpxA<sup>CA</sup>) is an essential domain for the hydrolysis of ATP and the binding of inhibitors and considered to be a promising target for broad-spectrum antimicrobial drugs development. The ATP-binding pocket in the CA domain contains a flexible ATP lid motif. Although the crystal structure of CA domain has been defined, the structure of the ATP lid remains uncertain, posing a challenge to the study of its catalytic mechanism. In this study, we report the backbone <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N chemical shift assignments of CpxA<sup>CA</sup> by heteronuclear multidimensional spectroscopy and predict its secondary structure in solution using TALOS<sup>+</sup>. The residues of ATP lid motif are well-assigned. Therefore, this study provides a foundation for understanding the role of CpxA<sup>CA</sup> in cellular signaling and the development of novel antimicrobial therapies.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143363515","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}
{"title":"Backbone and side‑chain <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments and secondary structure determination of the rhizobial FixJ.","authors":"Akio Horikawa, Rika Okubo, Naoki Hishikura, Riki Watanabe, Kaori Kurashima-Ito, Pooppadi Maxin Sayeesh, Kohsuke Inomata, Masaki Mishima, Hiroyasu Koteishi, Hitomi Sawai, Yoshitsugu Shiro, Teppei Ikeya, Yutaka Ito","doi":"10.1007/s12104-025-10221-w","DOIUrl":"https://doi.org/10.1007/s12104-025-10221-w","url":null,"abstract":"<p><p>The symbiotic nitrogen-fixing bacterium Bradyrhizobium japonicum (B.japonicum) enables high soybean yields with little or no nitrogen fertiliser. A two component regulatory system comprising FixL, a histidine kinase with O<sub>2</sub>-sensing activity, and FixJ, a response regulator, controls the expression of genes involved in nitrogen fixation, such as fixK and nifA. Only under anaerobic conditions, the monophosphate group is transferred from FixL to the N-terminal receiver domain of FixJ (FixJ<sub>N</sub>), which eventually promote the association of the C-terminal effector domain (FixJ<sub>C</sub>) to the promoter regions of the nitrogen-fixation-related genes. Structural biological analyses carried out so far for rhizobial FixJ molecules have proposed a solution structure for FixJ that differs from the crystal structures, in which the two domains are extended. To understand the FixJ activation caused by phosphorylation of the N-terminal domain, which presumably regulates through the interactions between FixJ<sub>N</sub> and FixJ<sub>C</sub>, here we have performed backbone and sidechain resonance assignments of the unphosphorylated state of B. japonicum FixJ.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143073384","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}
{"title":"<sup>1</sup>H, <sup>13</sup>C, <sup>15</sup>N and <sup>31</sup>P chemical shift assignment of the first stem-loop Guanidine-II riboswitch from Escherichia coli.","authors":"Tatjana Koob, Silas Döpp, Harald Schwalbe","doi":"10.1007/s12104-025-10217-6","DOIUrl":"https://doi.org/10.1007/s12104-025-10217-6","url":null,"abstract":"<p><p>A comprehensive understanding of RNA-based gene regulation is a fundamental aspect for the development of innovative therapeutic options in medicine and for a more targeted response to environmental problems. Within the different mechanisms of RNA-based gene regulation, riboswitches are particularly interesting as they change their structure in response to the interaction with a low molecular weight ligand, often a well-known metabolite. Four distinct classes of riboswitches recognize the very small guanidinium cation. We are focused on the Guanidine-II riboswitch with the mini-ykkC motif. We report here the assignment of the <sup>1</sup>H, <sup>13</sup>C, <sup>15</sup>N and <sup>31</sup>P chemical shifts of the 23 nucleotide-long sequence of the first stem-loop of the Guanidine-II riboswitch aptamer from Escherichia coli. Despite its small size, the assignment of the NMR signals of this RNA proved to be challenging as it has symmetrical base pairs and palindromic character.</p>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143073383","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}