Carrier Protein Research Articles

Mutagenesis to Orient Conjugation and Preserve Self-adjuvant Properties of Flagellin in Conjugates.

Bacterial flagellins are unique for their capacity to activate both the innate and the adaptive immune response through a Toll-like receptor 5 (TLR5) signaling cascade. Used as a carrier protein in conjugate vaccines it is crucial to preserve their self-adjuvant properties during the conjugation step. Considering the absence of cysteine in the Salmonella enterica flagellin FliC sequence, we have investigated the impact of five mutations (A2C, K180C, T240C, D251C and S306C) alone or in combination on TLR5 activation. The FliC mutated at the four positions K180C, T240C, D251C and S306C displayed much the same activity as native flagellin whether the cysteine residues were free or conjugated. These results pave the way for the preparation of self-adjuvanting conjugate vaccines based on cysteine-mutated FliC as a carrier protein.

The structure of full-length AFPK supports the ACP linker in a role that regulates iterative polyketide and fatty acid assembly

The polyketide synthases (PKSs) in microbes and the cytoplasmic fatty acid synthases in humans (FASs) are related enzymes that have been well studied. As a result, there is a paradigm explaining in general terms how FASs repeatedly use a set of enzymatic domains to produce simple fats, while PKSs use the domains in a much more complex manner to produce pharmaceuticals and other elaborate molecules. However, most animals also have PKSs that do not conform to the rules described in microbes, including a large family of enzymes that bridge fatty acid and polyketide metabolism, the animal FAS-like PKSs (AFPKs). Here, we present the cryoelectron microscopy structures of two AFPKs from sea slugs. While the AFPK resemble mammalian FASs, their chemical products mimic those of PKSs in complexity. How then does the architecture of AFPKs facilitate this structural complexity? Unexpectedly, chemical complexity is controlled not solely by the enzymatic domains but is aided by the dynamics of the acyl carrier protein (ACP), a shuttle that moves intermediates between these domains. We observed interactions between enzyme domains and the linker-ACP domain, which, when manipulated, altered the kinetic properties of the enzyme to change the resulting chemical products. This unveils elaborate mechanisms and enzyme motions underlying lipid and polyketide biochemistry across the domains of life.

Acinetobacter baumannii represses type VI secretion system through a manganese-dependent small RNA-mediated regulation.

Type VI secretion system (T6SS) is utilized by many Gram-negative bacteria to eliminate competing bacterial species and manipulate host cells. Acinetobacter baumannii ATCC 17978 utilizes T6SS at the expense of losing pAB3 plasmid to induce contact-dependent killing of competitor microbes, resulting in the loss of antibiotic resistance carried by pAB3. However, the regulatory network associated with T6SS in A. baumannii remains poorly understood. Here, we identified an Mn2+-dependent post-transcriptional regulation of T6SS mediated by a bonafide small RNA, AbsR28. A. baumannii utilizes MumT, an Mn2+-uptake inner membrane transporter, for the uptake of extracellular Mn2+ during oxidative stress. We demonstrate that the abundance of intracellular Mn2+ enables complementary base pairing of AbsR28-tssM mRNA (that translates to TssM, one of the vital inner membrane components of T6SS), inducing RNase E-mediated degradation of tssM mRNA and resulting in T6SS repression. Thus, AbsR28 mediates a crosstalk between MumT and T6SS in A. baumannii.IMPORTANCESmall RNAs (sRNAs) are identified as critical components within the bacterial regulatory networks involved in fine regulation of virulence-associated factors. The sRNA-mediated regulation of type VI secretion system (T6SS) in Acinetobacter baumannii was unchartered. Previously, it was demonstrated that A. baumannii ATCC 17978 cells switch from T6- to T6+ phenotype, resulting in the loss of antibiotic resistance conferred by plasmid pAB3. Furthermore, the derivatives of pAB3 found in recent clinical isolates of A. baumannii harbor expanded antibiotic resistance genes and multiple determinants for virulence factors. Hence, the loss of this plasmid for T6SS activity renders A. baumannii T6+ cells susceptible to antibiotics and compromises their virulence. Our findings show how A. baumannii tends to inactivate T6SS through an sRNA-mediated regulation that relies on Mn2+ and retains pAB3 during infection to retain antibiotic resistance genes carried on the plasmid.

Theranostic Near-Infrared Monoamine Oxidase Inhibitor (NMI) Protein Binding Interactions with MAOA and Albumin.

The protein binding interactions of near-infrared monoamine oxidase inhibitor (NMI) are reported here. NMI-bound proteins were examined by fluorescent SDS-PAGE and mass spectrometry using tumor tissues from brain and colon cancer mouse models. This study shows protein interactions with NMI, a chemical conjugate of MAOA inhibitor clorgyline and tumor-seeking dye, MHI-148. NMI fluorescence in MAOA knock-out (KO) mice was significantly lower compared to WT mice, including whole animal, organs, and tissue lysates which indicated that NMI binds to MAOA. Pure recombinant MAOA protein was detectable as a single fluorescent band that migrated at ~ 65kD. NMI inhibited MAOA activity (IC50 1-5µM). In a glioma mouse model, NMI targeted specifically to tumor with high contrast to adjacent normal brain, shown by a 65 kD protein band. Recent studies demonstrated heptamethine cyanine dyes (e.g., MHI-148) interact with serum albumin, contributing to tumor uptake and cancer cell internalization. Our study shows NMI binds to albumin but highly prefers MAOA, providing a plausible mechanism for systemic drug delivery via serum albumin to the tumor target and subsequent MAOA inhibition. Further studies in a colon cancer mouse model found the ~ 65 kD SDS-PAGE band, bound to NMI, contained both MAOA and albumin proteins by mass spectrometry. NMI was shown to interact with MAOA and the blood carrier protein, albumin. This study provides insights for drug delivery and protein target specificity of NMI to image and treat cancer.

Advancements in Isoniazid-Based Heterocyclic Derivatives as Potent Anti-Tubercular Agents: A Comprehensive Review of Synthesis, SAR and Biological activity (2017–2023)

Tuberculosis remains a significant global health issue that necessitates the synthesis of novel and innovative therapeutic agents. Isoniazid is a drug of choice for the treatment of tuberculosis, as it enables KatG to synthesize nicotinamide adenine dinucleotide. The enzyme reduction enzymes InhA and acyl carrier protein reductase are rendered inactive by these additives. This process generates type II fatty acids, which subsequently generate mycolic acid, a potent cell killing agent. Long-term, pervasive use—even misuse—is the primary cause of the increasing prevalence of INH-resistant mycobacterial strains. Numerous studies have demonstrated that heterocyclic scaffolds, when combined with isoniazid, exhibit exceptional anti-tubercular activity by enhancing the drug's penetration of bacterial cells. The review emphasizes the importance of heterocyclic moieties, such as phenylisoxazole, indanyl, indole, and isatin, in enhancing pharmacokinetic properties and overcoming drug resistance. We have concentrated on the isoniazid combined heterocyclic derivatives that were investigated as potential anti-tubercular agents from 2018 to 2023. This review aims to provide a valuable resource for researchers in the pursuit of more effective anti-tubercular therapies by guiding future research and development of isoniazid-based heterocyclic derivatives.

Membrane transport engineering for efficient yeast biomanufacturing.

Yeast strains have been widely recognized as useful cell factories for biomanufacturing. To improve production efficiency, their biosynthetic pathways and regulatory strategies have been continuously optimized. However, commercial production using yeasts is still limited by low product yield and high production cost. Accumulating evidences have demonstrated the importance of metabolite transport processes in addressing these challenges. Engineering yeast membrane transporters for transporting precursors, substrates, intermediates, products and toxic inhibitors has been successful. In addition, membrane properties are also important for metabolite production. Here we propose membrane transport engineering (MTE) to integrate manipulation of both membrane transporters and membrane properties. We emphasize that systematic optimization of both transporters and membrane lipid bilayers benefits production efficiency. We also envision the potential of artificial intelligence and automation process in MTE for economic and sustainable bioproduction using yeast cell factories.

Physicochemical features of subunit interfaces and their role in self-assembly across the ferritin superfamily.

Ferritins are ubiquitous and play a critical role in iron homeostasis. They are classified into four main subfamilies: classical, bacterial, bacterioferritin, and Dps. These are characterized by subunits with a four-helical bundle domain and interact through three distinct regions-one antiparallel interface (IntA) and two perpendicular interfaces (IntB and IntC), collectively forming a cage-like structure. Here, we attempt to characterize the variability of these interfaces across subfamilies. We found that IntA is essential for the dimeric unit assembly and is likely to assemble first, followed by the smaller interfaces of IntB and IntC (in any order), which are crucial for cage formation. These interfaces are unique in that they are less packed, although chemically stable, and their size lies between that of protein-protein complex and obligate homodimers. This study provides a detailed exploration of the ferritin interfaces, offering insights into their assembly and their importance as carrier proteins.

Dilated cardiomyopathy variant R14del increases phospholamban pentamer stability, blunting dynamic regulation of calcium.

The sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) is a membrane transporter that creates and maintains intracellular Ca2+ stores. In the heart, SERCA is regulated by an inhibitory interaction with the monomeric form of the transmembrane micropeptide phospholamban (PLB). PLB also forms avid homo-pentamers, and the dynamic exchange of PLB between pentamers and SERCA is an important determinant of cardiac responsiveness to exercise. Here, we investigated two naturally occurring pathogenic variants of PLB: a cysteine substitution of Arg9 (R9C) and an in-frame deletion of Arg14 (R14del). Both variants are associated with dilated cardiomyopathy. We previously showed that the R9C mutation causes disulfide crosslinking and hyperstabilization of pentamers. While the pathogenic mechanism of R14del is unclear, we hypothesized this mutation may also alter pentamer stability. Immunoblots revealed a significantly increased pentamer: monomer ratio for R14del-PLB compared to WT-PLB. We quantified homo-oligomerization and SERCA-binding in live cells using fluorescence resonance energy transfer (FRET) microscopy. R14del-PLB showed an increased affinity for homo-oligomerization and decreased binding affinity for SERCA compared to WT. The data suggest that, like R9C, the R14del mutation stabilizes PLB in pentamers, decreasing its ability to regulate SERCA. The R14del mutation reduced the rate of PLB unbinding from pentamers after transient elevations of Ca2+, limiting the recovery of PLB-SERCA complexes. A computational model predicted that hyperstabilization of PLB pentamers by R14del impairs the ability of cardiac Ca2+ handling to respond to changing heart rates between rest and exercise. We postulate that impaired responsiveness to physiological stress contributes to arrhythmogenesis in human carriers of the R14del mutation.

Compromised PPARγ Mediated Impaired Placental Glucose Transport via the PI3K/AKT Signaling Pathway is Associated with FGR.

Fetal growth restriction (FGR) is a condition in which a fetus cannot grow to its full potential during pregnancy. It is a leading cause of perinatal mortality and morbidity. However, the underlying etiology remains elusive. Here, we report that peroxisome proliferator-activated receptor γ (PPARγ) is inactivated in the trophoblasts of human placenta of FGR-complicated pregnancies. In the FGR placentas, p-PI3KTyr458 and p-AKTSer473 levels were also lowered. Additionally, there was a reduction in GLUT3 and GLUT4 levels in the cell membrane. Consistently, FGR patients showed decreased glucose concentrations in both the placenta and umbilical cord blood compared to that in normal pregnancy. In mouse models, deletion of PPARγ in trophoblasts and RUPP surgery successfully induced FGR and replicated these changes. Modulating PPARγ activity using rosiglitazone or GW9662 in BeWo cells resulted in the activation or inhibition of the PI3K/AKT signaling pathway, as well as the promotion or reduction of membrane translocation of GLUT3 and GLUT4, ultimately affecting glucose uptake in trophoblast cells. MK-2206 blocked these regulatory effects of rosiglitazone in BeWo cells. Furthermore, the administration of rosiglitazone encapsulated in placenta-targeting nanoparticles improved the growth and development of fetal mice in the RUPP group. In summary, PPARγ in trophoblast cells orchestrates the translocation of GLUT3 and GLUT4 to the cellular membrane via the PI3K/AKT signaling pathway, thereby regulating cellular glucose uptake and transport. Dysfunctions in this mechanism are strongly associated with FGR. Therefore, targeted activation of PPARγ in the placenta may be a potentially efficacious intrauterine intervention for FGR.

MXene-rGO nanocomposite based electrochemical immunosensor for detection of endosulfan - An organochlorine pesticide.

Endosulfan (Ed), a widely used organochlorine pesticide, is classified as a persistent organic pollutant (POP). Its long half-life, resistance to degradation, and bioaccumulation in the food chain contaminates soil, water, and air. Such widespread environmental damage triggers monitoring its levels for ensuring compliance with safety regulations and protecting public health. In the current work, Ed was chemically altered and coupled with a carrier protein to elicit an immunological response. The purified in-house generated antibodies against Ed (Ed-Ab) were characterized by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). MXene, a class of 2D inorganic compounds, is known to depict significant optoelectrical potential. Herein, we have synthesized a novel nanocomposite of MXene and reduced graphene oxide (rGO). For designing the MXene-rGO biosensor, Ed-Ab were combined with the nanocomposite post characterization by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDAX), X-ray photoelectron spectroscopy (XPS) and Fourier Transform infrared spectroscopy (FTIR). Using the differential pulse voltammetry (DPV), electrochemical parameters such as pH, temperature, scan rate and response time of the electrode were evaluated. The fabricated electro-immunosensor was employed for the detection of endosulfan wherein the limit of detection (LOD) for Ed was determined to be 0.497ppt with a linear range of 0.1ppt-1ppm. The composed electrode's working efficacy and sensitivity against similar cross-reactive pesticides was also determined. The MXene-rGO based nanocomposite depicted potential for determination of Ed traces in environmental samples.

Engineering an Escherichia coli surface display platform based on an autotransporter from Stenotrophomonas maltophilia: Autodisplay of enzymes with low to high molecular weight.

Surface display technology has garnered significant attention for preparing efficient whole cell catalysts, while reported carrier proteins still cannot meet the demand to display various passenger domains, especially for those with high molecular weight. This study demonstrates that the autotransporter of esterase Est7 (E7AT) from Stenotrophomonas maltophilia played a decisive role in its efficient surface display. Guided by the original signal peptide, the surface display ratio of Est7 was determined as 89.67 % with the total enzymatic activity of 16.67 U/mL, which was much higher than 4.60 U/mL and 5.70 U/mL for the signal peptides derived from pectolase B (pelB) and cholera toxin B (ctxB), respectively. Then, the E7AT unit was successfully developed to surface display proteins with varying molecular weight from 19.3 kDa to 117.9 kDa, showing a more effective autodisplay ability than adhesin involved in diffuse adherence (AIDA-I), ice nucleation proteins (InaK and InaP), and outer membrane proteins (lipoprotein/ompA, MltA-interacting protein A, and yiaT) systems. Additionally, a galactosidase (GAL) displayed by E7AT was employed to hydrolyze lactose, achieving a promising hydrolysis rate of 31.63 % in 2 h. The displayed GAL retained 63.24 % and 41.41 % activity in third and sixth batch, respectively, indicating the considerable potential of E7AT in developing efficient whole cell catalysts.

Development of Mycobacterium tuberculosis Enoyl Acyl Reductase (InhA) Inhibitors: A Mini-Review

This review article delves into the critical role of Enoyl acyl carrier protein reductase (InhA; ENR), a vital enzyme in the NADH-dependent acyl carrier protein reductase family, emphasizing its significance in fatty acid synthesis and, more specifically, the biosynthesis of mycolic acid. The primary objective of this literature review is to elucidate diverse scaffolds and their developmental progression targeting InhA inhibition, thereby disrupting mycolic acid biosynthesis. Various scaffolds, including thiourea, piperazine, thiadiazole, triazole, quinazoline, benzamide, rhodanine, benzoxazole, and pyridine, have been systematically explored for their potential as InhA inhibitors. Noteworthy findings highlight thiadiazole and triazole derivatives, demonstrating promising IC50 values within the nanomolar concentration range. The review offers comprehensive insights into InhA's structure, structure-activity relationships, and a detailed overview of distinct scaffolds as effective inhibitors of InhA.

Synthesis, Antibacterial Activity and In Silico Study of 1-(2-ethyl acetate)-2-styryl 5-nitroimidazole Derivatives

Background: For more than six decades, the use of metronidazole has been limited to anaerobic microorganisms. However, there are accounts of metronidazole derivatives exhibiting strong effectiveness against facultative anaerobic bacteria, suggesting that there may be another mechanism of action for metronidazole. Recent studies have shown that the enzyme FabH (β-ketoacyl-acyl carrier protein synthase III), responsible for the first step of fatty acid biosynthesis (FAB), is a promising target for nitroimidazole derivatives that can be used as an effective anti-infective agent. Objective: This study aimed to synthesize 1-(2-ethyl acetate)-2-styryl nitroimidazole derivatives and evaluate their in vitro and in silico antibacterial activity Methods: We synthesized 2-styryl 5-nitroimidazole derivatives by first condensing metronidazole with benzaldehydes and then carrying out an acetylation reaction. We evaluated the antimicrobial activity of the synthesized compounds against three Gram-positive bacterial strains (Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus agalactiae) and three Gram-negative bacterial strains (Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) using a two-fold serial dilution MTT (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assay. Results: Compounds 2 and 4 exhibited the highest level of antibacterial effectiveness, with minimum inhibitory concentrations (MIC) of 1.56 μg/mL against S. agalactiae and 3.13 μg/mL against P. aeruginosa. Compounds 2 and 4 also exhibited potent activity against K. pneumonia, with an MIC value of 6.25 μg/mL and 12.5 μg/mL, respectively. Molecular docking studies revealed that both compounds have favorable hydrophobic and electrostatic interactions with conserved residues in the binding site of the E. coli β-Ketoacyl-acyl carrier protein synthase III (FabH) complex. Conclusion:: Acetylation of 2-styryl-5-nitroimidazoles improved both their biological activity and binding interaction with the target protein

Lipid metabolism during seed germination of Pistacia chinensis and its response to gibberellic acid

Lipid metabolism may play a critical role in fueling seed germination, but the knowledge of lipid metabolism during germination is still ambiguous. Here, we hypothesize that gibberellic acid (GA) promotes germination by means of enhancing lipid mobilization in Chinese pistachio (Pistacia chinensis Bunge), a species belonging to Anacardiaceae with high oil content in its seeds. A multi-omics approach has been applied to measure lipid mobilization during seed germination, and to identify the key regulators involved in GA-mediated lipid metabolism. The results indicated that GA contents increased, while IAA, ABA and JA contents decreased during seed germination. GA3 increased significantly in the two germination stages (i.e. imbibition and radicle protrusion), and it was more abundant than GA1 and GA4. In addition, the relative content of most lipids decreased during germination, and the differentially changed metabolites were significantly enriched in lipid metabolic pathways based on KEGG analysis. WGCNA indicated that GA3 was correlated with more genes in lipid metabolic pathways. Transcriptomic analysis further revealed that differentially expressed genes (DEGs) related to fatty acid biosynthesis, glycerolipid metabolism, glycerophospholipid metabolism and starch and sucrose metabolism were upregulated under GA3 application, such as the acetyl-CoA carboxylase biotin carboxyl carrier protein (ACCB), fatty acyl-ACP thioesterase B (FATB), diacylglycerol acyltransferase (DGAT) and DEFECTIVE IN ANTHER DEHISCENCE 1 (DAD1). Therefore, our study supports the hypothesis that GA promotes seed germination in P. chinensis by enhancing lipid mobilization. This study proposes a novel mechanism of lipid responses to exogenous GA, which contributes to a deep understanding of germination of oleaginous seeds.

Expanding the scope of copper artificial metalloenzymes: A potential fluorinase?

Biocatalysts for fluorination are rare, and thus of great interest for artificial enzyme design. Biohybrid catalysts including Cu-based DNAzymes and dinucleotide catalysts can catalyse enantioselective electrophilic fluorination of β-ketoesters. Here we report the investigation of Cu-based artificial metalloenzymes as catalysts for electrophilic fluorination reactions. A library of artificial copper proteins was prepared by bioconjugation of bidentate and tridentate nitrogen ligands to cysteine variants of the Sterol Carrier Protein 2L (SCP-2L) and subsequent addition of Cu(II) salts. The resulting copper proteins were screened for activity for the fluorination of β-ketoesters using Selectfluor. Under aqueous acidic conditions it was observed that the designed catalysts did not outcompete the uncatalysed background reaction. This work highlights that careful consideration of substrate reactivity and background reactions is needed when considering potential reactions for artificial metalloenzyme catalysis.

Engineered IgG Fc-conjugation prolongs the half-life of florfenicol and alleviates pneumonia in mice

Small molecule drugs often exhibit short half-lives, requiring frequent administrations to maintain therapeutic concentrations over an extended period. To address this issue, the fragment crystallizable (Fc) region of IgG, known to prolong the half-life of antibodies via its interaction with the Fc neonatal receptor, was harnessed as a carrier protein to extend the half-life of a small molecule drug, florfenicol. Florfenicol, was chemically coupled to a recombinant Fc protein expressed using the eukaryotic expression system in HEK293cells. The Fc-florfenicol conjugate exhibited a substantially prolonged half-life of from 3.8 to 9.1h compared to unconjugated florfenicol and demonstrated excellent therapeutic properties in treating pneumonia in a mouse model. Our results, combined with the literature analysis on Fc-small molecule conjugates, show that Fc can substantially enhance the drug's half-life and suggest the potential for its use as a carrier in novel delivery systems.

Production of the recombinant human riboflavin transporters SLC52A1, 3 and functional assay in proteoliposomes.

Riboflavin, the FMN and FAD precursor, is a crucial vitamin in cell metabolism. Its adsorption and tissue distribution are mediated by tree membrane transporters namely RFVT1-3. Mutations of their genes are associated with Riboflavin Transporter Deficiency. Moreover, derangements of the level of these transporters have been found in several human cancers. To obtain a suitable experimental tool for studying the function of the single proteins, for testing the effect of pathological mutations and for validating predicted ligands as candidate drugs, we have set up a proteoliposome system harbouring the functional RFVT1 or RFVT3. RFVT proteins have been produced in E. coli and purified to the homogeneity by affinity chromatography. The purified proteins show an apparent molecular mass of 45.6 or 48.4 kDa, which are very close to the theoretical mass of RFVT1 or RFVT3, respectively. The purified transporters have been reconstituted into proteoliposomes using a methodology previously pointed out for RFVT2. The transport of riboflavin shows cooperative kinetics with K0.5 values of 0.86 or 1.13 μM and Hill coefficients of 1.19 or 1.3 for RFVT1 or RFVT3, respectively. The K0.5 data of both the transporters are similar the Km reported in intact cell studies. The transporters are inhibited by the riboflavin analogues FMN and lumiflavin in agreement with the molecular docking simulations.

Antibody-secreting cell repertoires hold high-affinity anti-rocuronium specificities that can induce anaphylaxis in vivo.

Neuromuscular blocking agents (NMBA) are muscle relaxants used to assist mechanical ventilation but lead in 1/10,000 anesthesia to severe acute hypersensitivity reactions i.e., anaphylaxis. Incidences vary between types of NMBAs. Rocuronium, a widely used non-depolarizing aminosteroid NMBA, induces among the highest anaphylaxis rates. Rocuronium-induced anaphylaxis is proposed to rely on pre-existing rocuronium-binding antibodies, but no such antibodies have ever been identified. Identify rocuronium-specific antibody repertoires from plasma cells or plasmablasts of rocuronium-immunized mice, and determine the affinities, structures and anaphylactogenic potential of these antibodies for rocuronium. Herein, we engrafted rocuronium onto carrier proteins allowing immunization of mice against rocuronium, screening for rocuronium-specific antibody responses, and sorting of rocuronium-specific plasma cells using droplet microfluidics coupled to single-cell antibody gene (VH and VL) sequencing. The two different repertoires of >500 VH-VL pairs were oligoclonal, comprised of three major clonal families, and displayed convergence. Expressed as human IgG1, these antibodies demonstrated subnanomolar affinities for rocuronium with families either monospecific for rocuronium or cross-reactive only for closely-related NMBAs. Expressed as human IgE, they triggered human mast cell and basophil activation, and severe passive systemic anaphylaxis in mice humanized for the IgE receptor FcεRI. Co-crystal structures between rocuronium and antibody representatives of three different VH-VL families revealed distinct interaction modes with, however, the ammonium group involved systematically in the binding interface. This work identifies the epitopes of antibody reactivity to rocuronium, demonstrates anaphylactogenic potential of anti-rocuronium IgE and establishes the first mouse model of NMBA anaphylaxis.

Waterlogging stress mechanism and membrane transporters in soybean (Glycine max (L.) Merr.).

An excess of water is more harmful to plant growth, root growth and the uniformity of the plant population than a water deficit. Water is a crucial factor in the three basic stages of soybean development: germination, emergence and flowering/seed filling. Waterlogging is one of the biggest constraints to crop production and productivity in India and can occur at any stage in soybean. However, seeds and seedlings are damaged by waterlogging resulting in a significant reduction in grain yield. Seed yield and growth are significantly correlated at the seedling stage. In addition, the plant is under constant pressure due to changing environmental conditions and has difficulty withstanding these harsh, unpredictable and difficult situations. Membrane transporters are essential and play fundamental roles during waterlogging thereby facilitating cellular homeostasis and gaseous exchange, which support plant growth and development. This review highlights the genetic basis and mechanism of waterlogging tolerance in soybean and the role of climate in influencing the genetic makeup of the crop, paving the way for further development of improved soybean varieties. Simultaneously, the article highlights membrane transporters' importance in water-mediated stress in soybeans.

Comprehensive analysis of the succinylome in Vero cells infected with peste des petits ruminants virus Nigeria 75/1 vaccine strain.

Peste des petits ruminants virus (PPRV) is currently the only member of the Morbillivirus caprinae species within the genus Morbillivirus of the family Paramyoxviridae. PPRV causes a highly contagious disease in small ruminants, especially goats and sheep. Succinylation is a newly identified and conserved modification and plays an important role in host cell response to pathogen infection. However, the extent and function of succinylation in Vero cells during PPRV infection remains unknown. In this study, a global profile of the succinylome in Vero cells infected with PPRV Nigeria 75/1 vaccine strain (PPRVvac) was performed by dimethylation labeling-based quantitative proteomics analysis. A total of 2633 succinylation sites derived from 823 proteins were quantified. The comparative analysis of differentially succinylated sites revealed that 228 down-regulated succinylation sites on 139 proteins and 44 up-regulated succinylation sites on 38 proteins were significantly modified in response to PPRVvac infection, seven succinylation motifs were identified. GO classification indicated that the differentially succinylated proteins (DSuPs) mainly participated in cellular respiration, biosynthetic process and transmembrane transporter activity. KEGG pathway analysis indicated that DSuPs were related to protein processing in the endoplasmic reticulum. Protein-protein interaction networks of the identified proteins provided further evidence that various ATP synthase subunits and carbon metabolism were modulated by succinylation, while the overlapped proteins between succinylation and acetylation are involved in glyoxylate and dicarboxylate metabolism. The findings of the present study provide the first report of the succinylome in Vero cells infected with PPRVvac and provided a foundation for investigating the role of succinylation alone and its overlap with acetylation in response to PPRVvac.