Pilus Protein Research Articles

Effect of Immunization of The Pili Protein 65.5 kDa Klebsiella pneumoniae on IFN-γ Levels of Spleen BALB/c Mice

Klebsiella pneumoniae is a Gram-negative bacterium that poses a threat to the global community. Currently, no vaccine for K. pneumoniae is licensed by the Food and Drug Administration (FDA). The delay in the manufacture of the K. pneumoniae vaccine was because many vaccine candidates failed at the clinical trial stage due to adverse cross-reactions. Pili can be used as a choice as a vaccine candidate. Pili K. pneumoniae is an immunogenic substance that triggers an immune response, one of which is the cytokine IFN-γ. Splenic splenocytes are the main source of IFN-γ-producing cells. The aim of this study is to determine the effect of immunization pili protein 65.5 kDa K. pneumoniae on IFN-γ levels from spleen BALB/c mice. There were 3 groups, K1 as control given PBS, K2 given pili protein 65.5 kDa + adjuvant, and K3 given adjuvant. IFN-γ was then measured by the ELISA method and analyzed by the ANOVA test. The results of measuring IFN-γ levels using One-way ANOVA showed that the total for all groups was 243.50 ± 43.7 with p < 0.05, the Post Hoc LSD test was continued. The Post Hoc test showed significant differences between K1 control and K2 groups, and between K1 and K3 groups, but not between K2 and K3 groups. It can be concluded that immunization with 65.5 kDa of pili protein does not affect the increase in IFN-γ levels in the spleen of BALB/c mice.

Infection by a multidrug-resistant Corynebacterium diphtheriae strain: prediction of virulence factors, CRISPR-Cas system analysis, and structural implications of mutations conferring rifampin resistance.

Cases of diphtheria, even in immunized individuals, are still reported in several parts of the world, including in Brazil. New outbreaks occur in Europe and other continents. In this context, studies on Corynebacterium diphtheriae infections are highly relevant, both for a better understanding of the pathogenesis of the disease and for controlling the circulation of clones and antimicrobial resistance genes. Here we present a case of cutaneous infection by multidrug-resistant Corynebacterium diphtheriae and provide its whole-genome sequencing. Genomic analysis revealed resistance genes, including tet(W), sul1, cmx, rpoB2, rbpA and mutation in rpoB. We performed phylogenetic analyzes and used the BRIG to compare the predicted resistance genes with those found in genomes from other significant isolates, including those associated with some outbreaks. Virulence factors such as spaD, srtBC, spaH, srtDE, surface-anchored pilus proteins (sapD), nonfimbrial adhesins (DIP0733, DIP1281, and DIP1621), embC and mptC (putatively involved in CdiLAM), sigA, dtxR and MdbA (putatively involved) in post-translational modification, were detected. We identified the CRISPR-Cas system in our isolate, which was classified as Type II-U based on the database and contains 15 spacers. This system functions as an adaptive immune mechanism. The strain was attributed to a new sequence type ST-928, and phylogenetic analysis confirmed that it was related to ST-634 of C. diphtheriae strains isolated in French Guiana and Brazil. In addition, since infections are not always reported, studies with the sequence data might be a way to complement and inform C. diphtheriae surveillance.

Identification of small molecule inhibitors of the Chloracidobacterium thermophilum type IV pilus protein PilB by ensemble virtual screening

Antivirulence strategy has been explored as an alternative to traditional antibiotic development. The bacterial type IV pilus is a virulence factor involved in host invasion and colonization in many antibiotic resistant pathogens. The PilB ATPase hydrolyzes ATP to drive the assembly of the pilus filament from pilin subunits. We evaluated Chloracidobacterium thermophilum PilB (CtPilB) as a model for structure-based virtual screening by molecular docking and molecular dynamics (MD) simulations. A hexameric structure of CtPilB was generated through homology modeling based on an existing crystal structure of a PilB from Geobacter metallireducens. Four representative structures were obtained from molecular dynamics simulations to examine the conformational plasticity of PilB and improve docking analyses by ensemble docking. Structural analyses after 1 μs of simulation revealed conformational changes in individual PilB subunits are dependent on ligand presence. Further, ensemble virtual screening of a library of 4234 compounds retrieved from the ZINC15 database identified five promising PilB inhibitors. Molecular docking and binding analyses using the four representative structures from MD simulations revealed that top-ranked compounds interact with multiple Walker A residues, one Asp-box residue, and one arginine finger, indicating these are key residues in inhibitor binding within the ATP binding pocket. The use of multiple conformations in molecular screening can provide greater insight into compound flexibility within receptor sites and better inform future drug development for therapeutics targeting the type IV pilus assembly ATPase.

Increased levels of IL-4 in the Spleen of BALB/c Mice after 65.5 kDa Pili Protein Klebsiella pneumoniae Immunization

Pneumonia is an infection of the lung parenchyma caused by Klebsiella pneumoniae, resulting in a high mortality rate of millions each year. To reduce these deaths, one potential solution is to create a vaccine that utilizes virulence factors of this microorganism, such as pili. During the early phase of infection, these antigens have a crucial role and can stimulate the production of memory b cells. These cells are activated by IL-4 (interleukin-4) in lymphoid organs, such as the spleen. Pathogen exposure such as virulence factors can stimulate the secretion of IL-4 in the spleen organ. This study aims to investigate the role of Klebsiella pneumoniae pili in this process. The research conducted from May to December 2023 at the Microbiology Laboratory, Faculty of Medicine, University of Jember was purely experimental, using Balb/c mice with IL-4 levels in the spleen organ as variables. The study used 15 mice, divided into control (K1), adjuvant (K2), and antigen (K3) groups. The research data were analyzed using non-parametric tests, specifically the Kruskal-Wallis and Post Hoc tests. The Kruskal-Wallis test revealed significant differences (p=0.003). In the Post Hoc test, a significant difference was found between the control and antigen groups (p=0.002). The results conclusively demonstrate that induction of Klebsiella pneumoniae pili protein 65.5 kDa significantly increases IL-4 levels in the spleen. Future studies should consider adding serum specimens to provide additional information.

Functional and biochemical characterisation of remote homologues of type IV pili proteins PilN and PilO in Helicobacter pylori.

Helicobacter pylori encodes homologues of PilM, PilN and PilO from bacteria with Type IV pili, where these proteins form a pilus alignment complex. Inactivation of pilO changes H. pylori motility in semi-solid media, suggesting a link to the chemosensory pathways or flagellar motor. Here, we showed that mutation of the pilO or pilN gene in H. pylori strain SS1 reduced the mean linear swimming speed in liquid media, implicating PilO and PilN in the function, or regulation of, the flagellar motor. We also demonstrated that the soluble variants of H. pylori PilN and PilO share common biochemical properties with their Type IV pili counterparts which suggests their adapted function in the bacterial flagellar motor may be similar to that in the Type IV pili.

Bacterial flagella hijack type IV pili proteins to control motility

Bacterial flagella and type IV pili (TFP) are surface appendages that enable motility and mechanosensing through distinct mechanisms. These structures were previously thought to have no components in common. Here, we report that TFP and some flagella share proteins PilO, PilN, and PilM, which we identified as part of the Helicobacter pylori flagellar motor. H. pylori mutants lacking PilO or PilN migrated better than wild type in semisolid agar because they continued swimming rather than aggregated into microcolonies, mimicking the TFP-regulated surface response. Like their TFP homologs, flagellar PilO/PilN heterodimers formed a peripheral cage that encircled the flagellar motor. These results indicate that PilO and PilN act similarly in flagella and TFP by differentially regulating motility and microcolony formation when bacteria encounter surfaces.

SARS-CoV-2 Spike Protein-Expressing Enterococcus for Oral Vaccination: Immunogenicity and Protection.

The declaration of the conclusion of the COVID-19 pandemic notwithstanding, coronavirus remains prevalent in circulation, and the potential emergence of novel variants of concern introduces the possibility of new outbreaks. Moreover, it is not clear how quickly and to what extent the effectiveness of vaccination will decline as the virus continues to mutate. One possible solution to combat the rapidly mutating coronavirus is the creation of safe vaccine platforms that can be rapidly adapted to deliver new, specific antigens in response to viral mutations. Recombinant probiotic microorganisms that can produce viral antigens by inserting specific viral DNA fragments into their genome show promise as a platform and vector for mucosal vaccine antigen delivery. The authors of this study have developed a convenient and universal technique for inserting the DNA sequences of pathogenic bacteria and viruses into the gene that encodes the pili protein of the probiotic strain E. faecium L3. The paper presents data on the immunogenic properties of two E. faecium L3 vaccine strains, which produce two different fragments of the coronavirus S1 protein, and provides an assessment of the protective efficacy of these oral vaccines against coronavirus infection in Syrian hamsters.

Increased levels of IL-10 in the spleen after induction of pili protein 65.5 kDa Klebsiella pneumoniae

Background: One of the gram-negative bacteria often found as the cause of nosocomial infections is Klebsiella pneumoniae. This bacterium has a mortality rate of 28.3% due to ESBL strains that cause resistance to several antibiotics and the absence of a vaccine as a preventive measure. Objective: To determine the level of IL-10 in the spleen after induction of the protein pili K. pneumoniae 65.5 kDa in mice. Method: This study used mice spleen samples from 21 mice aged six to eight weeks in an experimental investigation with a randomised posttest-only control group design. There are three groups, K1 as a PBS-given control, 65.5 kDa antigen protein pills + Freund’s adjuvants are given to K2, and Freund's adjuvants are given to K3. Interleukin-10 concentrations were measured using the ELISA and analysed by a one-way ANOVA assay. Result: The research showed significant differences in each group at IL-10 levels after administration of the pili protein K. pneumoniae using one-way ANOVA (p = 0.036). The treatment group had the highest average levels of IL-10, so there was an increased association between exposure to pili protein and IL-10 levels in rats. A significant increase in IL-10 levels was found in the treatment group compared to the control and adjuvant groups. Conclusion: Induction of protein pili Klebsiella pneumoniae 65.5 kDa increases IL-10 levels in spleen mice.

Mucosal Vaccines against Bacterial and Viral Pathogens

The mucosal membranes of the human body play a crucial role in the development, maintenance, and regulation of barrier functions and immune homeostasis, representing an integral component of the overall immune system. Mucosal vaccines elicit immune processes in the lymphoid tissue associated with the mucosal membranes. A critical objective of mucosal immunization is the identification of an antigen delivery vector capable of ensuring optimal vaccine efficacy. The authors of this article have conducted extensive research on the probiotic properties of enterococci over an extended period. They employ a safe and beneficial probiotic strain, Enterococcus faecium L3, as a delivery vector for vaccine antigens. Initially, the gene encoding the pathogenicity factor Bac, derived from group B streptococci (Streptococcus agalactiae), was successfully integrated into the genome of the probiotic strain E. faecium L3. Intravaginal, oral, and intranasal mucosal immunization methods utilizing the L3-Bac+ probiotic, which expresses antigenic determinants of pathogenic streptococci, were found to confer protection against bacterial infection in laboratory animals. Subsequently, recombinant technologies were refined, leading to the development of a universal method for incorporating a region of interest from the gene into the structure of the major pili protein gene of E. faecium L3. Using this technology, candidate vaccines against various infections, including Streptococcus pneumoniae, influenza A virus, and SARS-CoV-2 following the onset of the Covid-19 pandemic, have been obtained and tested. In this study, alongside the presentation of our own data, the challenges associated with utilizing recombinant probiotic bacteria as vectors for vaccine antigen delivery are discussed.

Mining of key genes for cold adaptation from Pseudomonas fragi D12 and analysis of its cold-adaptation mechanism.

The psychrotroph Pseudomonas fragi D12, which grew strongly under low temperatures, was screened from tundra soil collected from the permanent alpine zone on Changbai Mountain. To mine the genes critical for cold tolerance and to investigate the cold-adaptation mechanism, whole-genome sequencing, comparative genomic analysis, and transcriptome analysis were performed with P. fragi. A total of 124 potential cold adaptation genes were identified, including nineteen unique cold-adaptive genes were detected in the genome of P. fragi D12. Three unique genes associated with pili protein were significantly upregulated at different degrees of low temperature, which may be the key to the strong low-temperature adaptability of P. fragi D12. Meanwhile, we were pleasantly surprised to find that Pseudomonas fragi D12 exhibited different cold-adaptation mechanisms under different temperature changes. When the temperature declined from 30°C to 15°C, the response included maintenance of the fluidity of cell membranes, increased production of extracellular polymers, elevation in the content of compatibility solutes, and reduction in the content of reactive oxygen species, thereby providing a stable metabolic environment. When the temperature decreased from 15°C to 4°C, the response mainly included increases in the expression of molecular chaperones and transcription factors, enabling the bacteria to restore normal transcription and translation. The response mechanism of P. fragi D12 to low-temperature exposure is discussed. The results provide new ideas for the cold-adaptation mechanism of cold-tolerant microorganisms.

Engineering nanowires in bacteria to elucidate electron transport structural–functional relationships

Bacterial pilin nanowires are protein complexes, suggested to possess electroactive capabilities forming part of the cells’ bioenergetic programming. Their role is thought to be linked to facilitating electron transfer between cells and the external environment to permit metabolism and cell-to-cell communication. There is a significant debate, with varying hypotheses as to the nature of the proteins currently lying between type-IV pilin-based nanowires and polymerised cytochrome-based filaments. Importantly, to date, there is a very limited structure–function analysis of these structures within whole bacteria. In this work, we engineered Cupriavidus necator H16, a model autotrophic organism to express differing aromatic modifications of type-IV pilus proteins to establish structure–function relationships on conductivity and the effects this has on pili structure. This was achieved via a combination of high-resolution PeakForce tunnelling atomic force microscopy (PeakForce TUNA™) technology, alongside conventional electrochemical approaches enabling the elucidation of conductive nanowires emanating from whole bacterial cells. This work is the first example of functional type-IV pili protein nanowires produced under aerobic conditions using a Cupriavidus necator chassis. This work has far-reaching consequences in understanding the basis of bio-electrical communication between cells and with their external environment.

Front Cover

FEBS LettersVolume 597, Issue 10 p. 1317-1318 Issue InformationFree Access Front Cover First published: 24 May 2023 https://doi.org/10.1002/1873-3468.14388AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Graphical Abstract Cover illustration Clostridium perfringens minor/tip pilin (CppB) adopts an L-shaped structure in open form, and a small β-sheet unique to CppB provides a favorable binding site for collagen. The image refers to Tamai et al. ‘Structural and biochemical characterization of Clostridium perfringens pili protein B collagen-binding domains’. Volume597, Issue10May 2023Pages 1317-1318 RelatedInformation

Structural and biochemical characterization of Clostridium perfringens pili protein B collagen-binding domains.

Sortase-mediated pili are flexible rod proteins composed of major and minor/tip pilins, playing important roles in the initial adhesion of bacterial cells to host tissues. The pilus shaft is formed by covalent polymerization of major pilins, and the minor/tip pilin is covalently attached to the tip of the shaft involved in adhesion to the host cell. The Gram-positive bacterium Clostridium perfringens has a major pilin, and a minor/tip pilin (CppB) with the collagen-binding motif. Here, we report X-ray structures of CppB collagen-binding domains, collagen-binding assays and mutagenesis analysis, demonstrating that CppB collagen-binding domains adopt an L-shaped structure in open form, and that a small β-sheet unique to CppB provides a scaffold for a favourable binding site for collagen peptide.

Expanding strain coverage of a Group A Streptococcus pilus-expressing Lactococcus lactis mucosal vaccine.

Group A Streptococcus (GAS) is a human pathogenic bacterium that can trigger a wide range of diseases, including the autoimmune diseases acute rheumatic fever and rheumatic heart disease, causing major morbidity and mortality in many low- and middle-income countries. Primary intervention programs have had limited success thus far, and a licenced vaccine has yet to be developed. The pilus of GAS is known to be involved in host cell adhesion, biofilm formation and immune evasion. We have a mucosal vaccine in development that expresses the pilus of GAS on the surface of the non-pathogenic bacterium Lactococcus lactis. To expand strain coverage, we combined seven L. lactis constructs, each expressing a different GAS pilus variant and investigated the systemic and mucosal immune responses following immunisation. Mice immunised with this combination showed specific IgG and IgA responses to the GAS pilus proteins of vaccine strains, at levels comparable to mice immunised with a single construct. Cross-reactivity to pilus proteins of non-vaccine strains was also evident. Furthermore, protective efficacy against a homologous strain of GAS in a murine nasopharyngeal colonisation model was observed. Overall, this study provides further evidence for using pilus-expressing lactic acid bacteria as a vaccine to prevent upper respiratory tract GAS infections.

Method for creating a recombinant strain of enterococcus L3-SARS based on biologically active strain Enterococcus faecium L3

The current pandemic caused by the SARS-Cov-2 virus has significantly influenced the emergence of new injectable vaccines that provide a predominantly specific IgG response. However, it is generally accepted that protection against pathogens at the mucosal surface, which is the first barrier to viral entry, is predominantly dependent on the IgA response. It is now widely accepted that the use of genetically modified microorganisms, including probiotics, allows the oral or nasal mucosal delivery of therapeutic molecules, inducing an immune response in the mucous membranes. Probiotic strains are well studied for safety for the organism and are able to remain viable after passing through the gastric barrier, improve intraepithelial connections, and can generate a number of surface expressed molecules that enhance the effectiveness of vaccination.Recombinant probiotic microorganisms capable of producing vaccine antigens by inserting specific DNA fragments into their genome are one of the potential platforms that can be used to develop an appropriate vaccine containing a specific antigen for rapid response to viral mutations. Here, we demonstrate the construction of a novel SARS-Cov-2 vaccine candidate employing the gene fragment of S1 SARS-Cov-2 gene. According to the available data on new variants of SARS-Cov-2 mutations, three amino acid substitutions were made in the chosen sequence. This DNA fragment was inserted in frame into major pili protein gene within d2 domain of enterococcal operon encoding for pili.

Cj0683 Is a Competence Protein Essential for Efficient Initialization of DNA Uptake in Campylobacter jejuni.

C. jejuni is an important food-borne pathogen displaying high genetic diversity, substantially based on natural transformation. The mechanism of DNA uptake from the environment depends on a type II secretion/type IV pilus system, whose components are partially known. Here, we quantified DNA uptake in C. jejuni at the single cell level and observed median transport capacities of approximately 30 kb per uptake location. The process appeared to be limited by the initialization of DNA uptake, was finite, and, finalized within 30 min of contact to DNA. Mutants lacking either the outer membrane pore PilQ or the inner membrane channel ComEC were deficient in natural transformation. The periplasmic DNA binding protein ComE was negligible for DNA uptake, which is in contrast to its proposed function. Intriguingly, a mutant lacking the unique periplasmic protein Cj0683 displayed rare but fully functional DNA uptake events. We conclude that Cj0683 was essential for the efficient initialization of DNA uptake, consistent with the putative function as a competence pilus protein. Unravelling features important in natural transformation might lead to target identification, reducing the adaptive potential of pathogens.

Variable disruption of epithelial monolayers by Neisseria meningitidis carriage isolates of the hypervirulent MenW cc11 and MenY cc23 lineages.

Colonization of mucosal tissues by Neisseria meningitidis requires adhesion mediated by the type IV pilus and multiple outer-membrane proteins. Penetration of the mucosa and invasion of epithelial cells are thought to contribute to host persistence and invasive disease. Using Calu-3 cell monolayers grown at an air-liquid interface, we examined adhesion, invasion and monolayer disruption by carriage isolates of two clonal complexes of N. meningitidis. Carriage isolates of both the serogroup Y cc23 and the hypervirulent serogroup W cc11 lineages exhibited high levels of cellular adhesion, and a variable disruption phenotype across independent isolates. Inactivation of the gene encoding the main pilus sub-unit in multiple cc11 isolates abrogated both adhesive capacity and ability to disrupt epithelial monolayers. Contrastingly, inactivation of the phase-variable opa or nadA genes reduced adhesion and invasion, but not disruption of monolayer integrity. Adherence of tissue-disruptive meningococci correlated with loss of staining for the tight junction protein, occludin. Intriguingly, in a pilus-negative strain background, we observed compensatory ON switching of opa genes, which facilitated continued adhesion. We conclude that disruption of epithelial monolayers occurs in multiple meningococcal lineages but can vary during carriage and is intimately linked to pilus-mediated adhesion.

Peptide Sequence of Pili Subunit Protein 49.8 kDa Shigella flexneri as Antigenic Epitope for Shigellosis Vaccine Development.

This study investigates the amino acid sequence and identifies antigenic epitopes of 49.8 kilodalton (kDa) pili protein Shigella flexneri, which will be used as candidates for the shigellosis vaccine. Our study is a prospectively descriptive laboratory. We used bacterial isolate of S. flexneri pili isolation was performed using a pili cutter and sodium dodecyl-sulfate polyacrylamide gel electrophoresis. The amino acid sequences were analyzed using liquid chromatography dual mass spectrometry (LC-MS/MS) method in the proteomic laboratory. The target epitope antigenicity analysis was tested using Kolaskar and Tongaonkar Antigenicity software. The Bepired Linear Epitope Prediction software is used for epitope mapping. PymOL software was used for the visualization of proteins and molecular docking. Peptides and antibodies were applied to hemagglutination test and immune response was tested using the dot blot method. LC-MS/MS analysis results from the mascot server showed that the 49.8 kDa pili protein is S. flexneri similar to the flagellin protein of S. flexneri 1235-66 (ID I6H2T2). The results of antigenicity analysis and epitope mapping showed that areas of protein that has the most potential and antigenic epitopes are the regions 98-111 and 263-290 with the amino acid sequences, QSSTGTNSQSDLDS (Q-S) and DTTITKAETKTVTKNQVVDTPVTTDAAK (D-K). The results of the molecular docking interaction test between the peptide and the B-cell receptor have a low binding energy. Peptide Q-S and peptide D-K antigens are hemagglutinin molecules because they can agglutinate erythrocytes. The immune response between peptide antigens and anti-peptide antibodies can react based on color gradations in the dotblot method. The amino acid sequences Q-S and D-K are potentially antigenic epitopes. These peptides can be used to develop candidates for shigellosis vaccine.

Cryo-EM structure of the Agrobacterium tumefaciens T-pilus reveals the importance of positive charges in the lumen

Agrobacterium tumefaciens is a natural genetic engineer that transfers DNA into plants, which is the most applied process for generation of genetically modified plants. DNA transfer is mediated by a type IV secretion system in the cell envelope and extracellular T-pili. We here report the cryo-electron microscopic structures of the T-pilus at 3.2-Å resolution and of the plasmid pKM101-determined N-pilus at 3-Å resolution. Both pili contain a main pilus protein (VirB2 in A.tumefaciens, TraM in pKM101) and phospholipids arranged in a five-start helical assembly. They contain positively charged amino acids in the lumen, and the lipids are positively charged in the T-pilus (phosphatidylcholine) conferring overall positive charge. Mutagenesis of the lumen-exposed Arg91 in VirB2 results in protein destabilization and loss of pilus formation. Our results reveal that different phospholipids can be incorporated into type IV secretion pili and that the charge of the lumen may be of functional importance.

RETRACTED: Effect of Geobacter metallireducens nanowire on electron transfer efficiency in microbial fuel cell

AbstractBACKGROUNDTo understand the effect of electron shuttle compounds on the electron transfer behavior of MFC and the formation process of microbially fabricated nanowires. In this paper, 2,6‐anthraquinone desulfonation (AQDS) and Fe(OH)3 were used as electron mediator and electron acceptor, respectively, to study the effect of electron mediator on the formation of bio‐nanowires during the growth of Geobacter metallireducens in the double chamber MFC reactor (MFC), and the effects of electron transfer efficiency and electrical current characteristic.RESULTSThe results show that the culture process of the Geobacter metallireducens with Fe(OH)3 as an electron acceptor, the reduction product Fe(II) concentration in solution without AQDS was higher than that with AQDS after 10 days due to the formation of microbial nanowires. In comparison, the Pili protein content of the system without AQDS reached 336 ug/mL, which was higher than the AQDS system. Nanowires increased the transfer efficiency between biofilm and electrodes, which increased the maximum output voltage of MFC was 442 mV.CONCLUSIONAt the same time, it was found by electrochemical measurements that the nanowire biofilm electrode had a huge peak of the cyclic voltammetry curve, a small activation resistance, and a strong current response signal, indicating that the nanowires enhanced the electrochemical activity of the electrode. In addition, an electron transfer medium (AQDS) inhibited the expression of aromatic amino acids, tryptophan, and mtrC. Moreover, it destroyed the overlapping π‐π orbits of the aromatic parts, resulting in reduced and thinner nanowires and then decreasing the conductivity of synthetic organic materials like metals. © 2022 Society of Chemical Industry (SCI).