Analytical biochemistry最新文献

{"title":"Continuous assay for the dNTP triphosphohydrolase of activated SAMHD1","authors":"Roozbeh Eskandari,&nbsp;Daniel P. Groom,&nbsp;Ryo Tamura,&nbsp;Vern L. Schramm","doi":"10.1016/j.ab.2025.115966","DOIUrl":"10.1016/j.ab.2025.115966","url":null,"abstract":"<div><div>Sterile alpha motif and histidine-aspartate domain-containing protein 1 (SAMHD1) is the only member of the triphosphoric monoester hydrolase family in humans (dNTP + H₂O → dN + PPPi). The dNTPase activity of SAMHD1 inhibits DNA synthesis, resulting in cell-cycle arrest and restricting viral replication. The complex allosteric regulation mechanism of SAMHD1 and a reaction that lacks a direct spectroscopic signal make its kinetic analysis and inhibitor discovery challenging. We describe a continuous assay for monitoring SAMHD1 phosphatase activity in its activated physiological state. The assay uses a sequential assembly to generate the active tetrameric form of the enzyme. Two phosphatases convert inorganic triphosphate (PPPi) to inorganic phosphate (Pi). The released Pi reacts with the 7-methyl-6-thioguanosine and purine nucleoside phosphorylase to provide a sensitive continuous spectrophotometric assay. The assay is suitable for 96-microwell plate formats to provide a continuous measurement of SAMHD1 activity. The assay is benchmarked with inhibitors of SAMHD1. With a Z-prime value &gt; 0.90, the assay can be used for high-throughput screening of inhibitors for SAMHD1 and characterizing the allosteric or catalytic activity of the new inhibitors.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"708 ","pages":"Article 115966"},"PeriodicalIF":2.5,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144991184","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":"Revisiting phenylketonuria: Do high brain glycine levels caused by chronic hyperphenylalanemia contribute to brain dysfunction by modulating D-serine levels and NMDA receptor activity?","authors":"Gerald A. Dienel","doi":"10.1016/j.ab.2025.115992","DOIUrl":"10.1016/j.ab.2025.115992","url":null,"abstract":"<div><div>Phenylketonuria (PKU) is an inborn error of metabolism owing to deficits in phenylalanine hydroxylase (PAH) activity. PKU children acquire irreversible brain damage if newborns are not identified and treated with a phenylalanine-restricted diet. In spite of decades of research, the mechanisms underlying PKU brain dysfunction are not adequately understood. Competition of phenylalanine with large neutral amino acids (LNAAs) for carrier-mediated uptake into brain, causing lower brain LNAA levels and reduced neurotransmitter synthesis from tyrosine and tryptophan, is a long-favored mechanism for brain dysfunction. Here, glycine is hypothesized to contribute to phenylalanine-evoked brain disorders. All PKU animal models exhibit elevated brain glycine levels similar to mouse models of nonketotic hyperglycinemia. Glycine is synthesized from <span>l</span>-serine; it is a co-agonist of <em>N</em>-methyl-<span>d</span>-aspartate receptors (NMDARs) and an inhibitory neurotransmitter. <span>l</span>-Serine is synthesized from glucose in astrocytes, exported to neurons, and converted by serine racemase to <span>d</span>-serine, an NMDAR co-agonist. Increased glycine level enhances its inhibition of serine racemase and reduces levels of <span>d</span>-serine. <span>l</span>-Serine-glycine-<span>d</span>-serine interactions can be linked to PAH deficits because elevated brain phenylalanine concentration causes its metabolism by minor pathways to generate phenyllactate. If phenyllactate and <span>l</span>-serine synthesis are coupled via transaminase and redox reactions, the stoichiometry is 1:1. These findings support the following hypothesis: (i) phenylalanine disrupts glycine and <span>d</span>-serine homeostasis during brain maturation, irreversibly altering neuronal development and circuit formation, and (ii) high glycine and low <span>d</span>-serine levels in <span>PKU</span> adults contribute to cognitive and behavioral dysfunction. Suggested new directions for future studies of PKU focus on glycine neurotoxicity.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"708 ","pages":"Article 115992"},"PeriodicalIF":2.5,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145306838","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":"Mapping transposon insertion sites within bacterial genomes by direct Sanger sequencing","authors":"Scott W. Herke ,&nbsp;Linda M. Heffernan ,&nbsp;William N. Beavers ,&nbsp;Basel H. Abuaita","doi":"10.1016/j.ab.2025.115971","DOIUrl":"10.1016/j.ab.2025.115971","url":null,"abstract":"<div><div>Microbial biomedical research frequently involves mutagenesis by insertion of transposons into the genome. Currently, transposon insertion locations are typically elucidated by Next Generation Sequencing or by Sanger sequencing of PCR products. Here, transposons were located by direct Sanger sequencing of bacterial genomic DNA from both <em>Salmonella enterica</em> (Gram-negative) and <em>Staphylococcus aureus</em> (Gram-positive) cultures. DNA was prepared by shearing to a modal size of ∼2 kb followed by purification by paramagnetic beads. Sequencing reactions involved relatively minor modifications of standard protocols (e.g., extra sequencing polymerase, 75–100 PCR cycles); completed reactions were cleaned by ethanol-EDTA precipitation. Reads were generated on the ABI 3130xl Genetic Analyzer using 50-cm capillary arrays and a run protocol modified for extra sample injection time. Good quality reads of ∼500–800 nt were routinely generated; BLAST results returned nearly 100% matches to genomes in the NCBI database. As implemented, the optimized protocol (post-DNA extraction) could be performed within an 8-h workday (with sequencing results the following day) for ∼$10 (USD) per sequencing reaction. Although this method was developed to locate transposons inserted into bacterial genomes, it seems likely that it can be extended to generate sequence data from even native single-copy genes from small genomes (e.g., &lt;5 Mb).</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"708 ","pages":"Article 115971"},"PeriodicalIF":2.5,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145051605","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":"Differential binding of copper and zinc to a TDP-43 RNA recognition motif decapeptide and disulfide formation at residues C173/5 revealed by ESI-MS/MS","authors":"Josephine Esposto ,&nbsp;Naomi L. Stock ,&nbsp;Robert J. Huber ,&nbsp;Sanela Martic","doi":"10.1016/j.ab.2025.116031","DOIUrl":"10.1016/j.ab.2025.116031","url":null,"abstract":"<div><div>Copper (Cu) and zinc (Zn) metal ions play important roles in the proper functioning and localization of neurological proteins, such as transactive response DNA-binding protein 43 (TDP-43), which is linked to amyotrophic lateral sclerosis (ALS). Previous experimental and computational studies have identified putative Zn-binding regions within the RNA recognition motif 1 (RRM1) of TDP-43. However, Cu-binding interactions have been less explored despite their redox activity in regulating thiol (C173/175) conversion to disulfide within the RRM1 domain, influencing protein structure and function. Herein, the structural characterization and fragmentation pattern analysis of a TDP-43 decapeptide (166-HMIDGRWCDC-175), within RRM1, coordinated to Cu(II) and Zn(II) ions using electrospray ionization tandem mass spectrometry (ESI-MS/MS) was conducted under non-denaturing conditions. Higher-energy collision dissociation (HCD) fragmentation analysis identified that Cu(II) prefers His/Met residues, while Zn(II) was weakly coordinated to various binding sites in the peptide, specifically His, Met, Glu, Cys, Trp and Asp residues. Computational modeling using a metal ion binding server (MIB2) confirmed the binding sites and coordination sphere of metal-peptide complexes. No significant coordination to C173 and C175 was observed with Cu or Zn, as identified by using a double Cys mutant peptide. A complete thiol-to-disulfide conversion was observed in the presence of Cu(II)/(I) only, which was confirmed by the comparison of a preformed intramolecular disulfide peptide. Overall, unique differential coordination environments were observed for each metal ion with the peptide. The study provides new insights into metal ion interactions with TDP-43 RRM1 peptide, leading to a greater understanding of metal homeostasis in TDP-43 protein biochemistry and neurodegeneration.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"710 ","pages":"Article 116031"},"PeriodicalIF":2.5,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145762025","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":"Single-Entity Dual-Emissive MOF Platform for Reliable Ratiometric Point-of-Care Detection of Amoxicillin Residues","authors":"Sameera Sh. Mohammed Ameen ,&nbsp;Fiasal K. Algethami ,&nbsp;Khalid M. Omer ,&nbsp;Idrees B. Qader ,&nbsp;Hemn A. Qader","doi":"10.1016/j.ab.2025.116028","DOIUrl":"10.1016/j.ab.2025.116028","url":null,"abstract":"<div><div>In fluorescence-based sensing, self-referencing ratiometric analysis offers a significant advantage over external referencing by integrating both the probe and reference signals within a single material, rather than relying on two separate components. This intrinsic approach eliminates the need for additional reference dyes or materials, which can introduce inconsistencies due to variations in concentration, uneven dispersion, or environmental instability. Self-referencing materials provide a built-in correction mechanism, enhancing detection accuracy, reliability, and reproducibility while minimizing background interference. Despite their advantages, the design and synthesis of dual-emitting metal-organic frameworks (MOFs) with self-referencing capabilities remain rare and challenging. In this study, we introduce a novel Eu-based MOF with intrinsic dual-state, dual-emission properties, exhibiting distinct blue and red fluorescence in both liquid and solid states. The blue emission arises from the coordination-induced emission of the free, non-emissive ligand within the MOF structure. Interestingly, the red emission is selectively quenched by amoxicillin (AMX), while the blue fluorescence remains unaffected. This unique dual-emission feature enables a ratiometric sensing platform without requiring an external reference, ensuring greater stability, accuracy, and ease of use. With a linear detection range of 8.0–218 μM and a limit of detection of 0.354 μM, this Eu-MOF offers a robust and selective AMX sensing strategy. Additionally, a smartphone-assisted visual detection method using RGB analysis via the Color Grab App was developed, enabling portable and on-site quantification. This self-referencing Eu-MOF is inherently stable, recyclable, providing consistent signals and making it highly effective for pharmaceutical applications.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"710 ","pages":"Article 116028"},"PeriodicalIF":2.5,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145684009","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":"HCNS:A deep learning model for identifying essential proteins based on hypergraph convolution and sequence features","authors":"Jialong Tian ,&nbsp;Pengli Lu ,&nbsp;Huining Sha","doi":"10.1016/j.ab.2025.115949","DOIUrl":"10.1016/j.ab.2025.115949","url":null,"abstract":"<div><div>Accurate identification of essential proteins is crucial in biomedical research. Traditional methods rely on Protein–Protein Interaction (PPI) networks and limited biological feature data, often neglecting the critical role of protein amino acid sequences. To address this, we propose the HCNS model, which integrates the Hypergraph Convolutional Network (HGCN) module, the Seq-CNN-MB-NAG feature extraction module, and the Multi-Layer Perceptron (MLP) recognition module, significantly enhancing the accuracy of essential protein identification. The HGCN module constructs a hypergraph from the PPI network and protein complex data, capturing the complex structural relationships between proteins. The Seq-CNN-MB-NAG module utilizes Convolutional Neural Networks (CNN), Multi-Head Self-Attention (MHSA), Bidirectional Long Short-Term Memory (Bi-LSTM), and NAG Transformer to process protein amino acid sequences and extract sequence features. The MLP module then fuses these two sets of features for precise recognition. Experimental results show that the HCNS model outperforms existing methods, achieving an accuracy of 93.38%, with an Area Under the Curve (AUC) of 98.33% and an Area Under the Precision–Recall Curve (AUPR) of 97.16%, demonstrating its potential in essential protein identification.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"707 ","pages":"Article 115949"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144809717","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":"ITGB3 as a promising non-invasive biomarker for type 2 diabetes and diabetic nephropathy","authors":"Seyed Amirhossein Hosseini ,&nbsp;Parisa Ajorlou ,&nbsp;Hasti Haddadian ,&nbsp;Shahla Sohrabipour","doi":"10.1016/j.ab.2025.115965","DOIUrl":"10.1016/j.ab.2025.115965","url":null,"abstract":"<div><div>The prevalence of Type 2 diabetes mellitus (T2DM) is increasing worldwide and represents a major risk factor for the development of diabetic nephropathy (DN), a severe microvascular complication. Chronic hyperglycemia activates inflammatory and fibrotic signaling pathways, which contribute to kidney damage. Integrins, as transmembrane adhesion receptors, play pivotal roles in regulating inflammation, immune cell trafficking, and insulin resistance. This research focused on identifying non-invasive biomarkers for T2DM and DN using PBMCs. Differentially expressed genes related to diabetes were identified through the analysis of multiple datasets retrieved from the Gene Expression Omnibus, including <span><span>GSE95849</span><svg><path></path></svg></span>, <span><span>GSE9006</span><svg><path></path></svg></span>, <span><span>GSE25724</span><svg><path></path></svg></span>, and <span><span>GSE159984</span><svg><path></path></svg></span>. ITGB3 was identified as a common gene across these datasets, and its expression in DN was further examined using the <span><span>GSE142025</span><svg><path></path></svg></span> dataset. Real-time PCR analysis of PBMC samples revealed a significant upregulation of ITGB3 expression in individuals with DN and T2DM compared to healthy controls. The TF2DNA and miRNASNPv3 databases identified 10 transcription factors and 10 variants of ITGB3 involved in 60 miRNA interactions. Additionally, the DGIdb database revealed 15 drugs potentially regulating ITGB3 expression. These findings underscore the importance of integrin-related pathways in diabetes and suggest ITGB3 as a promising target for future research and therapeutic development.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"707 ","pages":"Article 115965"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144896119","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":"PreRBP: Interpretable deep learning for RNA-protein binding site prediction with attention mechanism","authors":"Huixian Chen ,&nbsp;Yun Zuo ,&nbsp;Xiangrong Liu ,&nbsp;Xiangxiang Zeng ,&nbsp;Zhaohong Deng ,&nbsp;Jiasong Wu","doi":"10.1016/j.ab.2025.115968","DOIUrl":"10.1016/j.ab.2025.115968","url":null,"abstract":"<div><div>In the complex process of gene expression and regulation, RNA-binding proteins occupy a pivotal position for RNA. Accurate prediction of RNA-protein binding sites can help researchers better understand RNA-binding proteins and their related mechanisms. And prediction techniques based on machine learning algorithms are both cost-effective and efficient in identifying these binding sites. However, there are some shortcomings in the currently available machine learning methods, such as the input features of the model only consider RNA sequence features, and most of the datasets suffer from class imbalance. To address these issues, this study first uses the publicly available 27 RNA-protein binding site datasets to construct a benchmark dataset. Then, we use RNAshapes and EDeN to obtain the secondary structure of RNA. Higher-order encoding method is used to extract the key information hidden in the RNA sequences and structures. In order to solve the class imbalance problem existing in the dataset, this study utilizes four undersampling algorithms, namely, random undersampling, NearMiss, ENN, and one-sided selection, to remove redundant samples in the negative samples, and lastly, based on Convolutional Neural Network, Bidirectional Long and Short Term Memory Network, this study constructs model PreRBP to predict RNA-protein binding sites.</div><div>The experimental results show that the model used in this study has an average AUC of 0.88, which is higher than other existing RNA-protein binding site prediction methods. Also, for the convenience of prediction, an online predictor is developed in this study. The predictor and experimental codes are available at <span><span>https://github.com/B12-Comet/RBPPrediction</span><svg><path></path></svg></span>.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"707 ","pages":"Article 115968"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145008102","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":"A novel dual-function sustainable method for H2O2 determination and dye biomineralization utilizing peroxidase-like MnFe2O4@CrFe2O4 nanocomposite","authors":"Saeed Reza Hormozi Jangi ,&nbsp;Masood Ranjoori ,&nbsp;Anahita Barghi ,&nbsp;Pardis Ahmadi","doi":"10.1016/j.ab.2025.115962","DOIUrl":"10.1016/j.ab.2025.115962","url":null,"abstract":"<div><div>In this contribution, a novel dual-function sustainable green nanozyme-mediated method for highly sensitive and selective H₂O₂ quantification and reusable rhodamine B biomineralization utilizing highly active MnFe₂O₄@CrFe₂O₄ nanocomposite with synergistic peroxidase-like activity was designed and developed. This method also introduced a sustainable approach for probing the analyte instead of exploiting prevalent carcinogenic nanozyme-based analytical probes, making it absolutely more sustainable than the conventional methods. The hydrogen peroxide biosensor acquired a linear range of 1–100 μM and a very low detection limit of 0.6 μM, along with an inter-day %RSD of 2.74 % and a highly selective response against coexisting materials. Ultimately, the sensor was employed for H₂O₂ quantification in milk, revealing a recovery of 96.1–102.8 %, %RSD = 1.4–3.6 %. Besides, the effective factors on decolorization yield were optimized, providing a high biomineralization yield of 99.4 % at optimal experimental conditions within a short time of 35.0 min. The breakthrough volume, storage stability, and reusability of the nanozymes were assessed, revealing a breakthrough volume of 5.0–1000 mL, a shelf-life of 20 days, and 70 % yield saving after 10 cycles. The method was applied for dye degradation in real water media, including river water, pool water, and tap water, revealing a high yield of over 95.4–99.5 %, %RSD = 1.8–4.2 %.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"707 ","pages":"Article 115962"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144893536","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":"A sensitive split luciferase assay for detecting olfactory receptor response to odorants","authors":"Natsumi Tanazawa ,&nbsp;Sho Obayashi ,&nbsp;Shuji Hinuma ,&nbsp;Shun'ichi Kuroda","doi":"10.1016/j.ab.2025.115954","DOIUrl":"10.1016/j.ab.2025.115954","url":null,"abstract":"<div><div>In this study, olfactory receptors (ORs) and Golf proteins fused with split luciferase were examined to develop a sensitive assay for identifying OR ligands. Although specific luminescence was observed from HEK293T cells expressing both fusion proteins (OR1A1-LgBiT and SmBiT-Mini-Golf) in response to an OR1A1 ligand, its intensity was low. To improve it, a fusion protein (V1N-G4S3-SmBiT-G4S3-Mini-Golf), was created where V1N contained the palmitoylation site necessary for localizing Mini-Golf to the cell membrane. When OR1A1-LgBiT and V1N-G4S3-SmBiT-G4S3-Mini-Golf were co-expressed in HEK293T cells, specific luminescence by ligand stimulation was considerably increased. The cell surface expression of OR1A1-LgBiT was markedly lower than that of OR1A1. Co-expression of receptor transporting protein 1 short form (RTP1S) promoted the cell surface expression of OR1A1-LgBiT more efficiently than OR1A1. Prediction using AlphaFold3 suggested that the N-terminal domain of RTP1S interacts with LgBiT in OR1A1-LgBiT. Co-expression of OR1A1-LgBiT, V1N-G4S3-SmBiT-G4S3-Mini-Golf, and RTP1S in HEK293T cells enhanced the sensitivity for detecting at least some ligands by 1000- to 3000-fold compared to the conventional cAMP assay. These findings suggest that optimizing the assembly of OR1A1-LgBiT and SmBiT-Mini-Golf at the cell membrane is essential for developing a sensitive assay. The OR response-detection system employing split luciferase holds significant potential for investigating ligands of orphan ORs.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"707 ","pages":"Article 115954"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144798011","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}