Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize adhesion of uropathogens; however, filament structure mechanism protection remain poorly understood. We used cryo-electron tomography to show uromodulin consists a zigzag-shaped backbone with laterally protruding arms. N-glycosylation mapping biophysical assays revealed acts as multivalent ligand for bacterial type 1 pilus adhesin, presenting specific epitopes on regularly spaced Imaging...

Abstract The overexpression of sialic acids on glycans, called hypersialylation, is a common alteration found in cancer cells. Sialylated glycans can enhance immune evasion by interacting with acid-binding immunoglobulin-like lectin (Siglec) receptors tumor-infiltrating Here, we investigated the effect sialylated and their interaction Siglec myeloid-derived suppressor cells (MDSCs). We that MDSCs derived from blood lung patients tumor-bearing mice strongly express inhibitory are highly...

N-Linked protein glycosylation is a frequent post-translational modification that can be found in all three domains of life. In canonical, highly conserved pathway, an oligosaccharide transferred by membrane-bound oligosaccharyltransferase from lipid donor to asparagines the sequon NX(S/T) secreted polypeptides. The δ-proteobacterium Actinobacillus pleuropneumoniae encodes unusual pathway for N-linked glycosylation. This takes place cytoplasm and mediated soluble N-glycosyltransferase (NGT)...

It is well known that mushrooms produce defense proteins and secondary metabolites against predators competitors; however, less about the correlation between tissue-specific expression target organism (antagonist) specificity of these molecules. In addition, conserved transcriptional circuitries involved in developing sexual organs fungi are not characterized, despite growing number gene datasets available from reproductive vegetative tissue. The aims this study were: first, to evaluate...

Abstract Bacteria are the main nutritional competitors of saprophytic fungi during colonization their ecological niches. This competition involves mutual secretion antimicrobials that kill or inhibit growth competitor. Over last years it has been demonstrated respond to presence bacteria with changes transcriptome, but significance these respect for nutrients is not clear as functional proof antibacterial activity induced gene products often lacking. Here, we report genome-wide...

The hallmark of N-linked protein glycosylation is the generation diverse glycan structures in secretory pathway. Dynamic, non-template-driven processes N-glycan remodeling endoplasmic reticulum and Golgi provide cellular setting for structural diversity. We applied newly developed mass spectrometry-based analytics to quantify site-specific model Pdi1p expressed insect cells. Molecular dynamics simulation, mutational analysis, kinetic studies vitro processing events flux analysis supported...

Abstract Exosomes are secreted into the extracellular space by most cell types and contain various molecular constituents, which play roles in many biological processes. Adipose‐derived mesenchymal stem cells (ADSCs) can differentiate a variety of secrete series paracrine factors through exosomes. ADSC‐derived exosomes have shown diagnostic therapeutic potential clinical diseases. The components critical for their mechanisms. Several methods been developed exosome purification, including...

The escalating menace of multidrug-resistant (MDR) pathogens necessitates a paradigm shift from conventional antibiotics to innovative alternatives. Antimicrobial peptides (AMPs) emerge as compelling contender in this arena. Employing silico methodologies, we can usher new era AMP discovery, streamlining the identification process vast candidate sequences, thereby optimizing laboratory screening expenditures. Here, unveil cutting-edge machine learning (ML) models that are both predictive and...

N-Linked protein glycosylation is a very common post-translational modification that can be found in all kingdoms of life. The classical, highly conserved pathway entails the assembly lipid-linked oligosaccharide and its transfer to an asparagine residue sequon NX(S/T) secreted by integral membrane oligosaccharyltransferase. A few species class γ-proteobacteria encode cytoplasmic N-glycosylation system mediated soluble N-glycosyltransferase (NGT). This enzyme uses nucleotide-activated sugars...

Kinetic analysis of intracellular intermediates affords a mathematical model for N-glycoprotein processing and secretion.

Equine chorionic gonadotropin (eCG) is a widely used hormone that synchronizes the female cycle and induces estrus in livestock. eCG heterodimeric glycoprotein composed of non-covalently linked α- β-chains whose glycosylation profiles determine vivo activity hormone. The commercially available products are crudely purified from serum pregnant mares, hence called mare (PMSG). Appropriate protein crucial for correct binding to receptor, receptor activation, its half-life. exact composition...

Duchenne muscular dystrophy (DMD) is a rare and severe genetic neuromuscular disease, characterized by rapid progression high mortality, highlighting the need for accurate ambulatory function assessment tools. Ultrasound imaging methods have been widely used quantitative analysis. Radiomics, which converts medical images into data, combined with machine learning (ML), offers promising solution. This study aimed at utilizing radiomics to analyze different stages of data generated during...

The inactivation of the ClC-0 chloride channel is very temperature sensitive and greatly facilitated by binding a zinc ion (Zn2+) from extracellular side, leading to Zn2+-induced current inhibition. To further explore relation Zn2+ inhibition inactivation, we mutated all 12 cysteine amino acids in assayed effect on these mutants. With this approach, found that C212 appears be important for sensitivity Upon mutating serine or alanine, macroscopic recordings disappears does not show detectable...

Abstract An expression map of the most abundant proteins in human hepatoma HepG2 cells was established by a combination complementary shotgun proteomics approaches. Two‐dimensional liquid chromatography (LC)‐nano electrospray ionization (ESI) tandem mass spectrometry (MS/MS) as well one‐dimensional LC‐matrix‐assisted laser desorption/ionization MS/MS were evaluated and shown that additional separation introduced at peptide level not efficient simple prefractionation protein extracts...

To study how the interaction between N-linked glycans and surrounding amino acids influences oligosaccharide processing, we used protein disulfide isomerase (PDI), a glycoprotein bearing 5 N-glycosylation sites, as model system expressed it transiently in Chinese hamster ovary (CHO)-S cell line. PDI was produced both secreted Sec-PDI endoplasmic reticulum-retained (ER)-PDI, to glycan processing by ER Golgi resident enzymes. Quantitative site-specific glycosylation profiles were obtained,...

The biosynthesis of eukaryotic lipid-linked oligosaccharides (LLOs) that act as donor substrates in protein N-glycosylation starts on the cytoplasmic side endoplasmic reticulum and includes sequential addition five mannose units to dolichol-pyrophosphate-GlcNAc2. These reactions are catalyzed by Alg1, Alg2 Alg11 gene products yield Dol-PP-GlcNAc2Man5, an LLO intermediate is subsequently flipped lumen reticulum. While purification active Alg1 has previously been described, have mostly studied...

Glycosylation of proteins profoundly impacts their physical and biological properties. Yet our ability to engineer novel glycoprotein structures remains limited. Established bacterial glycoengineering platforms require secretion the acceptor protein periplasmic space preassembly oligosaccharide substrate as a lipid-linked precursor, limiting access glycan substrates respectively. Here, we circumvent these bottlenecks by developing facile platform that operates in cytoplasm. The Glycoli...

Virus-like particles (VLPs) are emerging as nanoscaffolds in a variety of biomedical applications including delivery vaccine antigens and cargo such mRNA to mucosal surfaces. These soft, colloidal, proteinaceous structures (capsids) nevertheless susceptible environmental stress factors. We cross-linked multiple capsid surface amino acid residues using homobifunctional polyethylene glycol tethers improve the persistence survival model stressors. Surface cross-linking enhanced stability VLPs...

Significance Therapeutic antibodies represent the largest and fastest growing class of biopharmaceuticals. There is a trend in moving from intact toward “armed” antibody products, which moiety serves as pharmacodelivery vehicle. The impact glycosylation on targeting performance armed still largely unknown. Our article sheds light surprising finding that relatively small variations glycostructures sialic acid content can have dramatic effects therapeutic agent performance. A better...

N-Glycosylation is a post-translational modification performed in all three domains of life. In the halophilic archaea Haloferax volcanii, glycoproteins such as S-layer glycoprotein are modified by an N-linked pentasaccharide assembled series Agl (archaeal glycosylation) proteins. present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define structure this glycan attached at least four seven putative N-glycosylation sites, namely Asn-13, Asn-83,...

Atomistic glycoprotein simulations reveal a site-specific availability of glycan substrates in time-resolved mass spectrometry maturating enzyme kinetics.