Mina Yazdi, Tobias Burghardt, Johanna Seidl, Ulrich Lächelt, Ernst Wagner
{"title":"Evolution of Lipo-Xenopeptide Carriers for siRNA Delivery: Interplay of Stabilizing Subunits.","authors":"Mina Yazdi, Tobias Burghardt, Johanna Seidl, Ulrich Lächelt, Ernst Wagner","doi":"10.1021/acs.bioconjchem.5c00096","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00096","url":null,"abstract":"<p><p>Although small interfering RNA (siRNA) holds immense promise for treating genetic diseases and cancers, its clinical application is constrained by instability, cellular uptake barriers, and inefficient cytosolic delivery, underscoring the need for effective delivery systems. Therefore, this study focuses on screening novel T-shaped lipo-xenopeptide (XP) nanocarriers for siRNA polyplex formulation, integrating two single succinoyl-tetraethylene pentamine (Stp) units for electrostatic interaction and tyrosine tripeptides (Y<sub>3</sub>) for aromatic stabilization, along with structural modifications such as the addition of histidine (H) with or without terminal cysteines (C), and the incorporation of various fatty acids (FAs). A systematic evaluation of siRNA binding, nanoparticle stability, and gene silencing efficiency in multiple cell lines illustrated that the novel Stp1-HC lipo-XPs carriers outperform their Stp2-HC analogs, despite having fewer cationizable Stp units. This advantage stems from increased fatty acid, Y<sub>3</sub>, and C density, which compensates for reduced electrostatic interactions. The presence of H in combination with unsaturated FAs significantly improved the functional siRNA delivery. Our findings highlight the complex interplay of electrostatic, hydrophobic, covalent, hydrogen-bonded, and aromatic interactions to achieve efficient siRNA delivery, which is best-balanced in the oleic acid-containing Stp1-HC/OleA lipo-XP, exceeding the previously best standard carrier Stp2-HC/OleA in efficiency.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143707732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Valentina Greco, Valeria Lanza, Barbara Tomasello, Irina Naletova, Warren R L Cairns, Sebastiano Sciuto, Enrico Rizzarelli
{"title":"Copper Complexes with New Glycyl-l-histidyl-l-lysine-Hyaluronan Conjugates Show Antioxidant Properties and Osteogenic and Angiogenic Synergistic Effects.","authors":"Valentina Greco, Valeria Lanza, Barbara Tomasello, Irina Naletova, Warren R L Cairns, Sebastiano Sciuto, Enrico Rizzarelli","doi":"10.1021/acs.bioconjchem.4c00545","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00545","url":null,"abstract":"<p><p>In recent years, hyaluronic acid (HA) and the natural tripeptide glycyl-l-histidyl-l-lysine (GHK), especially its copper(II) complex (GHK-Cu), individually have been shown to exert helpful properties for bone protection and regeneration. However, they are not strong enough to handle oxidative stress, hydrolytic attack, or environmental conditions. Being aware that conjugation chemistry has recently emerged as an appealing approach for generating new molecular entities capable of preserving the molecular integrity of their moieties or delaying their degradation, herein we present the synthesis of conjugates of HA with GHK (GHK-HA), at different loadings of the tripeptide. GHK-HA binds copper(II) ions and potentiates the chemical and biological properties of the two components in in vitro assays. The results highlight copper's role in promoting the expression and release of certain trophic, angiogenic, and osteogenic factors, including brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), as well as bone morphogenetic protein-2 (BMP-2). The protective and regenerative activities of the metal ion are related to the translocation of its intracellular chaperones Copper Chaperone for Superoxide Dismutase (CCS) and Antioxidant-1 (Atox1) to the nucleus where they act as transcription factors.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143690465","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alva Abrahamsson, Andreas Berner, Justyna Golebiewska-Pikula, Namrata Chaudhari, Emelie Keskitalo, Cecilia Lindgren, Marcin K Chmielewski, Sjoerd Wanrooij, Erik Chorell
{"title":"Linker Design Principles for the Precision Targeting of Oncogenic G-Quadruplex DNA with G4-Ligand-Conjugated Oligonucleotides.","authors":"Alva Abrahamsson, Andreas Berner, Justyna Golebiewska-Pikula, Namrata Chaudhari, Emelie Keskitalo, Cecilia Lindgren, Marcin K Chmielewski, Sjoerd Wanrooij, Erik Chorell","doi":"10.1021/acs.bioconjchem.5c00008","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00008","url":null,"abstract":"<p><p>G-quadruplex (G4) DNA structures are noncanonical secondary structures found in key regulatory regions of the genome, including oncogenic promoters and telomeres. Small molecules, known as G4 ligands, capable of stabilizing G4s hold promise as chemical probes and therapeutic agents. Nevertheless, achieving precise specificity for individual G4 structures within the human genome remains a significant challenge. To address this, we expand upon G4-ligand-conjugated oligonucleotides (GL-Os), a modular platform combining the stabilizing properties of G4-ligands with the sequence specificity of guide DNA oligonucleotides. Central to this strategy is the linker that bridges the G4 ligand and the guide oligonucleotide. In this study, we develop multiple conjugation strategies for the GL-Os that enabled a systematic investigation of the linker in both chemical composition and length, enabling a thorough assessment of their impact on targeting oncogenic G4 DNA. Biophysical, biochemical, and computational evaluations revealed GL-Os with optimized linkers that exhibited enhanced binding to target G4s, even under thermal or structural stress. Notably, longer linkers broadened the range of targetable sequences without introducing steric hindrance, thereby enhancing the platform's applicability across diverse genomic contexts. These findings establish GL-Os as a robust and versatile tool for the selective targeting of individual G4s. By facilitating precise investigations of G4 biology, this work provides a foundation for advancing G4-targeted therapeutic strategies and exploring their role in disease contexts.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143668579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bioconjugate ChemistryPub Date : 2025-03-19Epub Date: 2025-02-24DOI: 10.1021/acs.bioconjchem.4c00580
Maia Marchand, Sébastien Depienne, Mohammed Bouzelha, Karine Pavageau, Roxane Peumery, Denis Loquet, Dimitri Alvarez-Dorta, Mickaël Guilbaud, Mikaël Croyal, Aurélien Dupont, Oumeya Adjali, Sébastien G Gouin, David Deniaud, Mathieu Mével
{"title":"Bioorthogonal Chemical Engineering of rAAV Capsid: Advancing Gene Therapy Targeting Using Proteins.","authors":"Maia Marchand, Sébastien Depienne, Mohammed Bouzelha, Karine Pavageau, Roxane Peumery, Denis Loquet, Dimitri Alvarez-Dorta, Mickaël Guilbaud, Mikaël Croyal, Aurélien Dupont, Oumeya Adjali, Sébastien G Gouin, David Deniaud, Mathieu Mével","doi":"10.1021/acs.bioconjchem.4c00580","DOIUrl":"10.1021/acs.bioconjchem.4c00580","url":null,"abstract":"<p><p>We report the chemical conjugation of a recombinant Adeno Associated Virus (rAAV) capsid with various functionalities, including proteins, using a bioorthogonal strategy. rAAVs were azido-coated or dibenzylcyclooctyne (DBCO)-coated by chemically modifying lysine or tyrosine residues. Lysine residues were modified using a phenyl isothiocyanate anchor, and tyrosine residues using either an aryl diazonium salt or a <i>N</i>-methyl luminol derivative. We demonstrate anchor-dependent labeling levels, as observed with biochemical assays and mass spectrometry. Strain-promoted azide-alkyne cycloaddition (SPAAC) was then implemented and evaluated on the rAAV to append functionalities such as fluorescein, biotin, and carbohydrates to the azido-coated capsids. We confirmed the efficiency of the bioorthogonal reaction and observed a stronger reactivity with dibenzylcyclooctyne (DBCO) compared to bicyclononyne (BCN). The optimized SPAAC reaction was finally used to label the viral vectors with two relevant nanobodies targeting specific immune cell receptors (CD62L and CD45). <i>In vitro</i> transduction assays conducted with one rAAV-nanobody conjugate demonstrated the promising targeting properties of these chemically modified vectors. Thus, we anticipate that this strategy will positively impact the field of rAAV capsid engineering and contribute in tissue-specific targeting for the optimization of gene therapy treatments.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"521-530"},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143490300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bioconjugate ChemistryPub Date : 2025-03-19Epub Date: 2025-02-27DOI: 10.1021/acs.bioconjchem.4c00589
Bryan Gosselin, Raphael Dutour, Julie Janssens, Ivan Jabin, Gilles Bruylants
{"title":"Repurposing Lateral Flow Assays as a Versatile and Rapid Characterization Tool for Bioconjugation of Nanoparticles.","authors":"Bryan Gosselin, Raphael Dutour, Julie Janssens, Ivan Jabin, Gilles Bruylants","doi":"10.1021/acs.bioconjchem.4c00589","DOIUrl":"10.1021/acs.bioconjchem.4c00589","url":null,"abstract":"<p><p>This study explores the use of lateral flow assays (LFAs), recognized for their simplicity and ease-of-use, as a tool for characterizing nanoparticles functionalized with various biomolecules (e.g., proteins, antibodies, and nucleic acids). A half-strip model system was developed using ovalbumin (OVA) conjugated to gold nanoparticles (AuNPs). The characterization results obtained with LFAs were compared to those from traditional methods such as infrared spectroscopy and fluorescence labeling. The advantages of LFAs in characterizing such conjugated nanosystems were clearly demonstrated. The use of half-strip assays could not only confirm the presence of OVA on AuNPs but also enable the quantification of OVA bound per nanoparticle, offering a rapid and quantitative characterization method. Additionally, the assay showcased its versatility, as it was successfully applied to optimize the covalent coupling conditions of OVA on AuNPs, as well as to differentiate between covalently bound and adsorbed proteins. Furthermore, LFAs were employed to detect antibodies on functionalized nanoparticles, optimize their coupling to a newly developed organic coating, and confirm both the grafting of nucleic acids onto the surface and their pairing with complementary strands. These findings underscore the remarkable adaptability of LFAs for characterizing diverse nanoconjugates. Overall, LFAs stand out as a versatile and accessible tool for characterizing complex bioconjugated nanosystems, making them highly suitable for rapid Quality Control (QC) analysis and bioconjugation optimization.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"531-539"},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143522200","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"<i>E. coli</i> as a Smart Thermo-Vector for Combating Solid Tumors: A Synergistic Heat-Induced Cancer Therapy Approach.","authors":"Tashmeen Kaur, Neeta Devi, Deepika Sharma","doi":"10.1021/acs.bioconjchem.5c00102","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00102","url":null,"abstract":"<p><p>Heat-induced cancer therapies such as magnetic hyperthermia-based cancer therapy (MHCT) and photothermal tumor ablation (PTT) have garnered significant attention as minimally invasive new-generation cancer therapy modalities. However, solid tumors associated with hypoxia present a considerable challenge to effective cancer therapy. In this study, we took up the challenge of mitigating the limiting penetration ability of nanoparticles by integrating polydopamine-coated magnetic nanoparticles and motile anaerobic bacteria (PDBs) to function as a smart thermo-vector. The developed PDBs are capable of self-navigating hypoxic tumors and as thermo-therapy agents with the ability to induce heat through exposure to an alternating magnetic field or near-infrared laser light. The thermo-vector system exhibited a dual-functioning synergistic antitumor effect of MHCT and PTT and an outstanding tumor targeting efficiency, outperforming the conventional 'nanoparticles only' approach. The heat-induced cellular oxidative stress and disrupted mitochondrial function led to 80% cellular cytotoxicity within 24 h of treatment. The PDB-based approach led to complete tumor regression in c57BL/6 mice within 21 days of treatment and a tumor-free survival for 60 days without recurrence, proving the capability of the developed PDBs in combatting solid tumors.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143655445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bioconjugate ChemistryPub Date : 2025-03-19Epub Date: 2025-02-17DOI: 10.1021/acs.bioconjchem.4c00541
Max S Kloet, Rishov Mukhopadhyay, Rukmini Mukherjee, Mohit Misra, Cami M P Talavera Ormeño, Rayman T N Tjokrodirijo, Paul J Hensbergen, Peter A van Veelen, Ivan Đikić, Aysegul Sapmaz, Gerbrand J van der Heden van Noort
{"title":"Comparison of Phosphoribosyl Ubiquitin Probes Targeting <i>Legionella</i> Dup Enzymes.","authors":"Max S Kloet, Rishov Mukhopadhyay, Rukmini Mukherjee, Mohit Misra, Cami M P Talavera Ormeño, Rayman T N Tjokrodirijo, Paul J Hensbergen, Peter A van Veelen, Ivan Đikić, Aysegul Sapmaz, Gerbrand J van der Heden van Noort","doi":"10.1021/acs.bioconjchem.4c00541","DOIUrl":"10.1021/acs.bioconjchem.4c00541","url":null,"abstract":"<p><p>In order to effectively replicate within a host cell, the <i>Legionella pneumophila</i> bacterium secretes effector enzymes into the cytoplasm in order to manipulate cellular host pathways including host ubiquitination. Some of these effectors, the so-called SidE-family, mediate noncanonical phosphoribosyl serine ubiquitination (PR-ubiquitination) of host substrate proteins, contributing to the recruitment of ER-remodeling proteins and the formation of a <i>Legionella</i>-containing vacuole, which is crucial in the early stages of bacterial infection. PR-ubiquitination is a dynamic process that is reversed by other <i>Legionella</i> effectors called deubiquitinases for PR-ubiquitination (Dups). We recently discovered a reactive allosteric cysteine in close proximity to the catalytic triad of DupA, which can be exploited as a target for covalent probe development. We here report on the synthesis of vinyl-sulfonate and fluoro-sulfonate warhead-containing phosphoribosyl ubiquitin probes, where the Arg42 position of ubiquitin is linked to the C1 of ribose via a native guanidinium group, and compare them to triazole-linked probes. In vitro tests on recombinant DupA and SdeA<sub>PDE</sub> revealed that these probes are able to capture the enzymes covalently. In a pull-down proteomics experiment, DupA and DupB enzymes are enriched from <i>Legionella</i>-infected cell lysates, highlighting the potential of native Arg-riboside linked probes to capture <i>Legionella</i> effector enzymes in a complex proteome.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"457-463"},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11926780/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bioconjugate ChemistryPub Date : 2025-03-19Epub Date: 2025-03-01DOI: 10.1021/acs.bioconjchem.4c00512
Seobin Kim, Jiyeon Kang, Danbi An, Jinho Seo, Doo-Byoung Oh
{"title":"Lysosome-Targeting Chimera Using Mannose-6-Phosphate Glycans Derived from Glyco-Engineered Yeast.","authors":"Seobin Kim, Jiyeon Kang, Danbi An, Jinho Seo, Doo-Byoung Oh","doi":"10.1021/acs.bioconjchem.4c00512","DOIUrl":"10.1021/acs.bioconjchem.4c00512","url":null,"abstract":"<p><p>Lysosome-targeting chimeras (LYTACs) harness the cell's lysosomal degradation machinery to break down extracellular and membrane proteins. Previous methods used a synthetic glycopeptide containing multiple serine-O-mannose-6-phosphate (poly-M6Pn), which presented challenges such as synthetic complexity and potential immunogenicity associated with poly-M6Pn. This study introduced a LYTAC formulation, LYTAC<sup>gyM6pG</sup>, which uses glyco-engineered yeast-derived mannose-6-phosphate glycans (gyM6pGs) for lysosomal transport, overcoming synthetic complexities and immunogenic risks. The gyM6pGs used in LYTAC<sup>gyM6pG</sup> are human-compatible (identical to the structures found in humans) and are efficiently produced through yeast fermentation, followed by the preparation of cell wall glycans and their <i>in vitro</i> modifications. We employed copper-free click chemistry (azide and dibenzocyclooctyne reactions) for the robust conjugation of gyM6pGs to a nanobody targeting the immune checkpoint protein PD-L1, thereby streamlining the assembly of LYTAC<sup>gyM6pG</sup>. We demonstrated that LYTAC<sup>gyM6pG</sup> effectively degraded endogenous and recombinant PD-L1 proteins on the cell surface by directing them to the lysosome via the cation-independent mannose-6-phosphate receptor pathway. Furthermore, LYTAC<sup>gyM6pG</sup> significantly enhanced T cell-mediated cytotoxicity against cancer cells, surpassing the efficacy of nanobodies alone. The successful application of gyM6pGs in the development of LYTAC<sup>gyM6pG</sup> highlights the potential for a more viable and scalable therapeutic production of LYTACs, paving the way for broader therapeutic applications, including cancer treatment.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"424-436"},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143530904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bioconjugate ChemistryPub Date : 2025-03-19Epub Date: 2025-03-05DOI: 10.1021/acs.bioconjchem.4c00567
Sandra Pinzón Martín, Jasmin Mecinović
{"title":"Selenalysine as a Chemical Tool for Probing Histone Post-Translational Modifications.","authors":"Sandra Pinzón Martín, Jasmin Mecinović","doi":"10.1021/acs.bioconjchem.4c00567","DOIUrl":"10.1021/acs.bioconjchem.4c00567","url":null,"abstract":"<p><p>Post-translational modifications (PTMs) on histones play a crucial role in determining the structure and function of chromatin, thereby regulating the eukaryotic gene expression. Histone lysine methylation and acetylation are among the most widespread and biomedically important PTMs, with new chemical tools for their examination in high demand. Here, we report the first use of γ-selenalysine as an efficient lysine mimic for enzymatic methylation, acetylation, and deacetylation reactions catalyzed by histone lysine methyltransferases, acetyltransferases, and a deacetylase. We also show that easily accessible selenocysteine and cysteine residues can undergo chemo- and site-selective alkylation reactions to generate both unmodified and modified γ-selenalysine and related γ-thialysine residues in histone peptides. This dual-modification strategy enables the site-specific incorporation of two distinct functionalities into peptides, mimicking lysine post-translational modifications commonly found on histones. Our research presents a novel approach in which selenocysteine serves as a unique handle for the chemoselective introduction of selenalysine, along with its methylated and acetylated analogues. These tools are designed to facilitate the study of epigenetic proteins that are important for human health and disease.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"510-520"},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143555306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bioconjugate ChemistryPub Date : 2025-03-19Epub Date: 2025-03-03DOI: 10.1021/acs.bioconjchem.4c00493
Md Shahadat Hossain, Md Mahbubul Alam, Zhiwei Huang, Faeze Mousazadeh, Ronit Sarangi, Ebbing de Jong, Kavindu C Kolamunna, Albert L Adhya, James L Hougland, Atanu Acharya, Davoud Mozhdehi
{"title":"Scalable One-Pot Production of Geranylgeranylated Proteins in Engineered Prokaryotes.","authors":"Md Shahadat Hossain, Md Mahbubul Alam, Zhiwei Huang, Faeze Mousazadeh, Ronit Sarangi, Ebbing de Jong, Kavindu C Kolamunna, Albert L Adhya, James L Hougland, Atanu Acharya, Davoud Mozhdehi","doi":"10.1021/acs.bioconjchem.4c00493","DOIUrl":"10.1021/acs.bioconjchem.4c00493","url":null,"abstract":"<p><p>Geranylgeranylation is a critical post-translational modification essential for various cellular functions. However, current methods for synthesizing geranylgeranylated proteins are complex and costly, which hinders access to these proteins for both biophysical and biomaterials applications. Here, we present a method for the one-pot production of geranylgeranylated proteins in <i>Escherichia coli</i>. We engineered <i>E. coli</i> to express geranylgeranyl pyrophosphate synthase (GGS), an enzyme that catalyzes the production of geranylgeranyl pyrophosphate. By coexpressing GGS with a geranylgeranyltransferase, we achieved efficient geranylgeranylation of model protein substrates, including intrinsically disordered elastin-like polypeptides (ELPs) and globular proteins such as mCherry and the small GTPases RhoA and Rap1B. We examined the biophysical behavior of the resulting geranylgeranylated proteins and observed that this modification affects the phase-separation and nanoassembly of ELPs and lipid bilayer engagement of mCherry. Taken together, our method offers a scalable, versatile, and cost-effective strategy for producing geranylgeranylated proteins, paving the way for advances in biochemical research, therapeutic development, and biomaterial engineering.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"415-423"},"PeriodicalIF":4.0,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11926785/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143539491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}