Structure, sequon recognition and mechanism of tryptophan C-mannosyltransferase.

C-linked glycosylation is essential for the trafficking, folding and function of secretory and transmembrane proteins involved in cellular communication processes. The tryptophan C-mannosyltransferase (CMT) enzymes that install the modification attach a mannose to the first tryptophan of WxxW/C sequons in nascent polypeptide chains by an unknown mechanism. Here, we report cryogenic-electron microscopy structures of Caenorhabditis elegans CMT in four key states: apo, acceptor peptide-bound, donor-substrate analog-bound and as a trapped ternary complex with both peptide and a donor-substrate mimic bound. The structures indicate how the C-mannosylation sequon is recognized by this CMT and its paralogs, and how sequon binding triggers conformational activation of the donor substrate: a process relevant to all glycosyltransferase C superfamily enzymes. Our structural data further indicate that the CMTs adopt an unprecedented electrophilic aromatic substitution mechanism to enable the C-glycosylation of proteins. These results afford opportunities for understanding human disease and therapeutic targeting of specific CMT paralogs.

Oxidative desulfurization pathway for complete catabolism of sulfoquinovose by bacteria.

Catabolism of sulfoquinovose (SQ; 6-deoxy-6-sulfoglucose), the ubiquitous sulfosugar produced by photosynthetic organisms, is an important component of the biogeochemical carbon and sulfur cycles. Here, we describe a pathway for SQ degradation that involves oxidative desulfurization to release sulfite and enable utilization of the entire carbon skeleton of the sugar to support the growth of the plant pathogen Agrobacterium tumefaciens SQ or its glycoside sulfoquinovosyl glycerol are imported into the cell by an ATP-binding cassette transporter system with an associated SQ binding protein. A sulfoquinovosidase hydrolyzes the SQ glycoside and the liberated SQ is acted on by a flavin mononucleotide-dependent sulfoquinovose monooxygenase, in concert with an NADH-dependent flavin reductase, to release sulfite and 6-oxo-glucose. An NAD(P)H-dependent oxidoreductase reduces the 6-oxo-glucose to glucose, enabling entry into primary metabolic pathways. Structural and biochemical studies provide detailed insights into the recognition of key metabolites by proteins in this pathway. Bioinformatic analyses reveal that the sulfoquinovose monooxygenase pathway is distributed across Alpha- and Betaproteobacteria and is especially prevalent within the Rhizobiales order. This strategy for SQ catabolism is distinct from previously described pathways because it enables the complete utilization of all carbons within SQ by a single organism with concomitant production of inorganic sulfite.

Yeast- and antibody-based tools for studying tryptophan C-mannosylation.

Tryptophan C-mannosylation is an unusual co-translational protein modification performed by metazoans and apicomplexan protists. The prevalence and biological functions of this modification are poorly understood, with progress in the field hampered by a dearth of convenient tools for installing and detecting the modification. Here, we engineer a yeast system to produce a diverse array of proteins with and without tryptophan C-mannosylation and interrogate the modification's influence on protein stability and function. This system also enabled mutagenesis studies to identify residues of the glycosyltransferase and its protein substrates that are crucial for catalysis. The collection of modified proteins accrued during this work facilitated the generation and thorough characterization of monoclonal antibodies against tryptophan C-mannosylation. These antibodies empowered proteomic analyses of the brain C-glycome by enriching for peptides possessing tryptophan C-mannosylation. This study revealed many new modification sites on proteins throughout the secretory pathway with both conventional and non-canonical consensus sequences.

Structural basis of substrate recognition and catalysis by fucosyltransferase 8.

Fucosylation of the innermost GlcNAc of N-glycans by fucosyltransferase 8 (FUT8) is an important step in the maturation of complex and hybrid N-glycans. This simple modification can dramatically affect the activities and half-lives of glycoproteins, effects that are relevant to understanding the invasiveness of some cancers, development of mAb therapeutics, and the etiology of a congenital glycosylation disorder. The acceptor substrate preferences of FUT8 are well-characterized and provide a framework for understanding N-glycan maturation in the Golgi; however, the structural basis of these substrate preferences and the mechanism through which catalysis is achieved remain unknown. Here we describe several structures of mouse and human FUT8 in the apo state and in complex with GDP, a mimic of the donor substrate, and with a glycopeptide acceptor substrate at 1.80-2.50 Å resolution. These structures provide insights into a unique conformational change associated with donor substrate binding, common strategies employed by fucosyltransferases to coordinate GDP, features that define acceptor substrate preferences, and a likely mechanism for enzyme catalysis. Together with molecular dynamics simulations, the structures also revealed how FUT8 dimerization plays an important role in defining the acceptor substrate-binding site. Collectively, this information significantly builds on our understanding of the core fucosylation process.

Synthesis of C-Mannosylated Glycopeptides Enabled by Ni-Catalyzed Photoreductive Cross-Coupling Reactions.

The biological functions of tryptophan C-mannosylation are poorly understood, in part, due to a dearth of methods for preparing pure glycopeptides and glycoproteins with this modification. To address this issue, efficient and scalable methods are required for installing this protein modification. Here, we describe unique Ni-catalyzed cross-coupling conditions that utilize photocatalysis or a Hantzsch ester photoreductant to couple glycosyl halides with (hetero)aryl bromides, thereby enabling the α-C-mannosylation of 2-bromo-tryptophan, peptides thereof, and (hetero)aryl bromides more generally. We also report that 2-(α-d-mannopyranosyl)-L-tryptophan undergoes facile anomerization in the presence of acid: something that must be considered when preparing and handling peptides with this modification. These developments enabled the first automated solid-phase peptide syntheses of C-mannosylated glycopeptides, which we used to map the epitope of an antibody, as well as providing the first verified synthesis of Carmo-HrTH-I, a C-mannosylated insect hormone. To complement this approach, we also performed late-stage tryptophan C-mannosylation on a diverse array of peptides, demonstrating the broad scope and utility of this methodology for preparing glycopeptides.

Photoredox-Catalyzed Generation of Sulfamyl Radicals: Sulfonamidation of Enol Silyl Ether with Chlorosulfonamide.

A novel and practical photoredox-catalyzed generation of sulfamyl radicals followed by radical sulfonamidation of enol silyl ether has been described. Diverse functionalized ß-ketosulfonamides were prepared in modest to excellent yields under mild and economic reaction conditions through the present catalytic protocol. Furthermore, the methodology developed provides an efficient and convenient approach to the synthesis of the antiseizure drug Zonisamide.

Generation of Aryl Radicals from Aryl Halides: Rongalite-Promoted Transition-Metal-Free Arylation.

A new and practical method for the generation of aryl radicals from aryl halides is reported. Rongalite as a novel precursor of super electron donors was used to initiate a series of electron-catalyzed reactions under mild conditions. These transition-metal-free radical chain reactions enable the efficient formation of C-C, C-S, and C-P bonds through homolytic aromatic substitution or SRN1 reactions. Moreover, the synthesis of antipsychotic drug Quetiapine was performed on gram scale through the described method. This protocol demonstrated its potential as a promising arylation method in organic synthesis.

N-Radical-Initiated Cyclization through Insertion of Sulfur Dioxide under Photoinduced Catalyst-Free Conditions.

A N-radical-initiated cyclization involving the sulfonylation of unactivated alkenes through insertion of sulfur dioxide in the presence of visible light under catalyst-free conditions is accomplished. A range of sulfonated 3,4-dihydro-2H-pyrroles can be generated in good yields under photoinduced sulfonylative conditions. Additionally, the corresponding 2-(3,4-dihydro-2H-pyrrol-2-yl)methylsulfonyl-1-arylethanones can be easily converted to 3,4-dihydro-2H-1,4-thiazine 1,1-dioxides. This photoinduced transformation occurs efficiently at room temperature under catalyst-free conditions, and various functional groups can be tolerated. A tandem radical process is involved through the iminyl radical-mediated cyclization with the insertion of sulfur dioxide; this process shows high efficiency and good selectivity.

Yttrium and Niobium Elements Co-Doping and the Formation of Double Perovskite Structure Ba2YNbO6 in BCZT.

The (Ba0.85Ca0.15) (Ti0.90Zr0.10)O3 + x Y3+ + x Nb5+ (abbreviated as BCZT-x(Nb + Y), x = 0 mol%, 0.05 mol%, 0.1 mol%, 0.2 mol%, 0.3 mol%) lead-free piezoceramics samples were prepared by a traditional solid-state sintering method. And the effects of Yttrium and Niobium elements (Y3+ and Nb5+) co-doping on the defect, phase and structure, microstructure, and comprehensive electrical properties have been investigated. Research results show that the Y and Nb elements co-doping can dramatically enhance piezoelectric properties. It is worth noting that XPS defect chemistry analysis, XRD phase analysis and TEM results together show that a new phase of double perovskite structure Barium Yttrium Niobium Oxide (Ba2YNbO6) is formed in the ceramic, and the XRD Rietveld refinement and TEM results show the coexistence of the R-O-T phase. Both these two reasons together lead to significant performance improvements of piezoelectric constant (d33) and planar electro-mechanical coupling coefficient (kp). The functional relation between temperature and dielectric constant testing results present that the Curie temperature increases slightly, which shows the same law as the change of piezoelectric properties. The ceramic sample reaches an optimal performance at x = 0.1% of BCZT-x(Nb + Y), where d33 = 667 pC/N, kp = 0.58, εr = 5656, tanδ = 0.022, Pr = 12.8 µC/cm2, EC = 2.17 kV/cm, TC =92 °C, respectively. Therefore, they can be used as potential alternative materials to lead based piezoelectric ceramics.

A water-stable dual-responsive Cd-CP for fluorometric recognition of hypochlorite and acetylacetone in aqueous media.

One novel cadmium(II)-coordination polymer [Cd3L2(datrz)(H2O)3] (CP 1) is controllably synthesized by surmising the astute combination of semi-rigid tricarboxylate acid 4-(2',3'-dicarboxylphenoxy) benzoic acid (H3L) and auxiliary ligand 3,5-diamino-1,2,4-triazole (datrz). Structure analysis shows that CP 1 has a two-dimensional (2D) layer structure with a 5-nodal (43) (44·62) (45·64·8) (45·6) (47·66·82) topology. Further investigations reveal that CP 1 shows superordinary water stability and good thermal stability. The fluorescent explorations suggest that the as-synthesized CP 1 could emit blue light centered at 485 nm, attributing to ligand-based emission. In terms of sensing investigations, CP 1 could act as a fluorescent sensor for detecting hypochlorite (ClO-) and acetylacetone (acac) through fluorescence turn-off process in aqueous solution, and the detection limit could reach 0.18 µM and 0.056 µM, respectively. Further research reveals that it is more likely the N-H···O-Cl hydrogen bonds between -NH2 groups of the triazole ligands and O atoms of ClO- plays the key role in the system, which may serve as a bridge for the energy transfer, leading to fluorescence quenching of the chemosensor. While the photoinduced electron transfer (PET) combined with inner filter effect (IFT) should be responsible for the turn-off fluorescence of CP 1 triggered by acac.

Influences of intergrowth structure construction on the structural and electrical properties of the BBT-BiT ceramics.

Bismuth Layer Structured Ferroelectrics (BLSFs) have always been an important research direction of high Curie temperature piezoelectrical ceramics, and the construction of intergrowth structure has been considered as an effective method to improve the electric properties of BLSFs. There are many literatures about intergrowth structure improving electrical performance, but few reports analyze the influence of the construction of intergrowth structure on the internal defects and electrical properties in BLSFs. In this study, (1-x) BaBi4Ti4O15 - x Bi4Ti3O12 ceramic samples with intergrowth bismuth layer structure were fabricated by a conventional solid-state reaction method, and the mechanism of the influence of intergrowth structure construction on the structure and electrical properties of BLSFs has been discussed. The crystal structure, phase composition, microstructure, dielectric and piezoelectric performance, relaxation behavior and AC conductivity of ceramic samples were systematically investigated. It has been found that the construction of intergrowth structure can significantly inhibit the generation of oxygen vacancies. The concentration of the oxygen vacancies plays an important role, and its reduction will lead to the inhibition of grain growth and the increase of the relaxation activation energy of ceramics. In addition, the intergrowth structure construction also affects the symmetry of ceramics in the c-axis direction, thus affecting the electrical properties of ceramics.

Motion Artifact Reduction In Photoplethysmography For Reliable Signal Selection.

Photoplethysmography (PPG) is a non-invasive and economical technique to extract vital signs of the human body. Although it has been widely used in consumer and research grade wrist devices to track a user's physiology, the PPG signal is very sensitive to motion which can corrupt the signal's quality. Existing Motion Artifact (MA) reduction techniques have been developed and evaluated using either synthetic noisy signals or signals collected during high-intensity activities - both of which are difficult to generalize for real-life scenarios. Therefore, it is valuable to collect realistic PPG signals while performing Activities of Daily Living (ADL) to develop practical signal denoising and analysis methods. In this work, we propose an automatic pseudo clean PPG generation process for reliable PPG signal selection. For each noisy PPG segment, the corresponding pseudo clean PPG reduces the MAs and contains rich temporal details depicting cardiac features. Our experimental results show that 71% of the pseudo clean PPG collected from ADL can be considered as high quality segment where the derived MAE of heart rate and respiration rate are 1.46 BPM and 3.93 BrPM, respectively. Therefore, our proposed method can determine the reliability of the raw noisy PPG by considering quality of the corresponding pseudo clean PPG signal.

Late Stage Phosphotyrosine Mimetic Functionalization of Peptides Employing Metallaphotoredox Catalysis.

Access to phosphotyrosine (pTyr) mimetics requires multistep syntheses, and therefore late stage incorporation of these mimetics into peptides is not feasible. Here, we develop and employ metallaphotoredox catalysis using 4-halogenated phenylalanine to afford a variety of protected pTyr mimetics in one step. This methodology was shown to be tolerant of common protecting groups and applicable to the late stage pTyr mimetic modification of protected and unprotected peptides, and peptides of biological relevance.

N-Radical Initiated Aminosulfonylation of Unactivated C(sp3)-H Bond through Insertion of Sulfur Dioxide.

N-Radical initiated aminosulfonylation of unactivated C(sp3)-H bond through insertion of sulfur dioxide in the presence of visible light is reported. O-Aryl oximes react with DABCO·(SO2)2 smoothly at room temperature under blue LED irradiation without any metals or photoredox catalysts, generating diverse 5,6-dihydro-4H-1,2-thiazine 1,1-dioxides in good yield. Additionally, this approach can be extended to the synthesis of 1H-benzo[d][1,2]thiazine 2,2-dioxides. During the reaction process, an N-radical is initiated by the treatment of O-aryl oximes with DABCO·(SO2)2 under visible-light irradiation. It is followed by aminosulfonylation of a nearby C(sp3)-H bond through 1,5-hydrogen atom transfer with accompanying insertion of sulfur dioxide to provide 1,2-thiazine 1,1-dioxide derivatives.