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1.
Plant J ; 117(3): 892-908, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-37955978

RESUMEN

Tetraspanins (TETs) are small transmembrane scaffold proteins that distribute proteins into highly organized microdomains, consisting of adaptors and signaling proteins, which play important roles in various biological events. In plants, understanding of tetraspanin is limited to the Arabidopsis TET genes' expression pattern and their function in leaf and root growth. Here, we comprehensively analyzed all rice tetraspanin (OsTET) family members, including their gene expression pattern, protein topology, and subcellular localization. We found that the core domain of OsTETs is conserved and shares a similar topology of four membrane-spanning domains with animal and plant TETs. OsTET genes are partially overlapping expressed in diverse tissue domains in vegetative and reproductive organs. OsTET proteins preferentially targeted the endoplasmic reticulum. Mutation analysis showed that OsTET5, OsTET6, OsTET9, and OsTET10 regulated plant height and tillering, and that OsTET13 controlled root growth in association with the jasmonic acid pathway. In summary, our work provides systematic new insights into the function of OsTETs in rice growth and development, and the data provides valuable resources for future research.


Asunto(s)
Arabidopsis , Oryza , Animales , Oryza/genética , Oryza/metabolismo , Tetraspaninas/genética , Tetraspaninas/metabolismo , Proteínas de la Membrana/metabolismo , Plantas/metabolismo , Arabidopsis/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas
2.
Nat Chem Biol ; 19(1): 101-110, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36229683

RESUMEN

Adenine base editors (ABEs) catalyze A-to-G transitions showing broad applications, but their bystander mutations and off-target editing effects raise safety concerns. Through structure-guided engineering, we found ABE8e with an N108Q mutation reduced both adenine and cytosine bystander editing, and introduction of an additional L145T mutation (ABE9), further refined the editing window to 1-2 nucleotides with eliminated cytosine editing. Importantly, ABE9 induced very minimal RNA and undetectable Cas9-independent DNA off-target effects, which mainly installed desired single A-to-G conversion in mouse and rat embryos to efficiently generate disease models. Moreover, ABE9 accurately edited the A5 position of the protospacer sequence in pathogenic homopolymeric adenosine sites (up to 342.5-fold precision over ABE8e) and was further confirmed through a library of guide RNA-target sequence pairs. Owing to the minimized editing window, ABE9 could further broaden the targeting scope for precise correction of pathogenic single-nucleotide variants when fused to Cas9 variants with expanded protospacer adjacent motif compatibility. bpNLS, bipartite nuclear localization signals.


Asunto(s)
Adenina , Edición Génica , Animales , Ratones , Ratas , Mutación , Citosina , Sistemas CRISPR-Cas/genética , ARN Guía de Sistemas CRISPR-Cas
3.
Eur J Neurosci ; 60(4): 4536-4551, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38978299

RESUMEN

During development, embryos and foetuses may be exposed to maternally ingested antiseizure medications (ASM), valproate and lamotrigine, essential in some patients to control their epilepsy symptoms. Often, the two drugs are co-administered to reduce required doses of valproate, a known potential teratogen. This study used Genetic Absence Epilepsy Rat from Strasbourg to evaluate transfer of valproate and lamotrigine across late gestation placenta and their entry into cerebrospinal fluid (CSF) and brain of developing rats, in mono- and combination therapies. Animals at embryonic day (E) 19, postnatal day (P) 0, 4 and 21, and adults were administered valproate (30 mg/kg) or lamotrigine (6 mg/kg) with their respective [3H]-tracers, either alone or in combination. In chronic experiments, females consumed valproate-containing diet from 2 weeks prior to mating until offspring were used at E19 and P0. Drugs were injected 30 min before blood, CSF and brain samples were collected from terminally anaesthetised animals. Radioactivity in samples was measured. In acute monotherapy brain entry of valproate was higher in foetal than postnatal animals, correlating with its plasma protein binding. Brain entry of lamotrigine was not age-dependent. Combination therapy enhanced entry of lamotrigine into the adult brain but had no effects on brain and CSF entry of valproate. Following chronic valproate exposure, placental transfer of valproate decreased in combination therapy; however, foetal brain entry increased. Results suggest that during pregnancy, the use of combination therapy of valproate and lamotrigine may mitigate overall foetal exposure to valproate but potential risks to foetal brain development are less clear.


Asunto(s)
Anticonvulsivantes , Encéfalo , Epilepsia Tipo Ausencia , Lamotrigina , Placenta , Triazinas , Ácido Valproico , Animales , Femenino , Embarazo , Anticonvulsivantes/administración & dosificación , Epilepsia Tipo Ausencia/tratamiento farmacológico , Epilepsia Tipo Ausencia/genética , Epilepsia Tipo Ausencia/metabolismo , Ratas , Placenta/metabolismo , Placenta/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/efectos de los fármacos , Triazinas/administración & dosificación , Intercambio Materno-Fetal , Masculino
4.
Anal Chem ; 96(36): 14320-14325, 2024 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-39208257

RESUMEN

Bioluminescence imaging (BLI) is an important noninvasive optical imaging technique that has been widely used to monitor many biological processes due to its high sensitivity, resolution, and signal-to-noise ratio. However, the BLI technique based on the firefly luciferin-luciferase system is limited by the expression of exogenous luciferase and the short half-life of firefly luciferin, which pose challenges for long-term tracking in vivo. To solve the problems, here we rationally designed an intelligent strategy for persistent BLI in tumors by combining luciferase-loaded calcium phosphate nanoparticles (Luc@CaP NPs) to provide luciferase and the probe Cys(SEt)-Lys-CBT (CKCBT) to slowly produce the luciferase substrate amino luciferin (Am-luciferin). Luc@CaP NPs constructed with CaP as a carrier could enable luciferase activity to be maintained in vivo for at least 12 h. And compared to the conventional substrate luciferin, CKCBT apparently prolonged the BL time by up to 2 h through GSH-induced intracellular self-assembly and subsequent protease degradation-induced release of Am-luciferin. We anticipate that this strategy could be applied for clinical translation in more disease diagnosis and treatment in the near future.


Asunto(s)
Neoplasias de la Mama , Fosfatos de Calcio , Luciferasas , Mediciones Luminiscentes , Nanopartículas , Fosfatos de Calcio/química , Nanopartículas/química , Animales , Mediciones Luminiscentes/métodos , Humanos , Luciferasas/metabolismo , Luciferasas/química , Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/patología , Femenino , Ratones , Imagen Óptica , Ratones Endogámicos BALB C , Línea Celular Tumoral , Ratones Desnudos
5.
Small ; 20(31): e2311351, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38453673

RESUMEN

Supramolecular self-assembly has emerged as an efficient tool to construct well-organized nanostructures for biomedical applications by small organic molecules. However, the physicochemical properties of self-assembled nanoarchitectures are greatly influenced by their morphologies, mechanical properties, and working mechanisms, making it challenging to design and screen ideal building blocks. Herein, using a biocompatible firefly-sourced click reaction between the cyano group of 2-cyano-benzothiazole (CBT) and the 1,2-aminothiol group of cysteine (Cys), an amino-acid-encoded supramolecular self-assembly platform Cys(SEt)-X-CBT (X represents any amino acid) is developed to incorporate both covalent and noncovalent interactions for building diverse morphologies of nanostructures with bioinspired response mechanism, providing a convenient and rapid strategy to construct site-specific nanocarriers for drug delivery, cell imaging, and enzyme encapsulation. Additionally, it is worth noting that the biodegradation of Cys(SEt)-X-CBT generated nanocarriers can be easily tracked via bioluminescence imaging. By caging either the thiol or amino groups in Cys with other stimulus-responsive sites and modifying X with probes or drugs, a variety of multi-morphological and multifunctional nanomedicines can be readily prepared for a wide range of biomedical applications.


Asunto(s)
Portadores de Fármacos , Nanoestructuras , Nanoestructuras/química , Portadores de Fármacos/química , Aminoácidos/química , Humanos , Cisteína/química
6.
Small ; : e2402763, 2024 Aug 25.
Artículo en Inglés | MEDLINE | ID: mdl-39183531

RESUMEN

The primary challenges in tumor imaging and therapy revolve around improving targeting efficiency, enhancing probe/drug delivery efficacy, and minimizing off-target signals and toxicity. Although various carriers have been developed, many are difficult to synthesize, costly, and not universally applicable. Furthermore, numerous carriers exhibit limited delivery rates in solid tumors, particularly larger nanocarriers. To address these challenges, a simple binary co-assembly drug delivery platform has been designed using the readily synthesized small molecule Cys(SEt)-Lys-CBT (CKCBT) as the self-assembly building block. CKCBT can effectively penetrate tumor cells due to its positively charged Lys side chain and small size. Upon glutathione reduction, CKCBT co-assembles with Nile red or Chlorin e6 to form nanofibers inside tumor cells. This enables their specific accumulation in tumor cells rather than normal cells and extends their exposure time, resulting in precise and enhanced tumor imaging and treatment. Hence, this uncomplicated and highly efficient binary co-assembly drug delivery platform can be easily adapted to a broad spectrum of probes and drugs, presenting a novel approach for advancing clinical diagnosis and therapy.

7.
Nat Methods ; 18(11): 1304-1316, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34725484

RESUMEN

Glycoproteomics is a powerful yet analytically challenging research tool. Software packages aiding the interpretation of complex glycopeptide tandem mass spectra have appeared, but their relative performance remains untested. Conducted through the HUPO Human Glycoproteomics Initiative, this community study, comprising both developers and users of glycoproteomics software, evaluates solutions for system-wide glycopeptide analysis. The same mass spectrometrybased glycoproteomics datasets from human serum were shared with participants and the relative team performance for N- and O-glycopeptide data analysis was comprehensively established by orthogonal performance tests. Although the results were variable, several high-performance glycoproteomics informatics strategies were identified. Deep analysis of the data revealed key performance-associated search parameters and led to recommendations for improved 'high-coverage' and 'high-accuracy' glycoproteomics search solutions. This study concludes that diverse software packages for comprehensive glycopeptide data analysis exist, points to several high-performance search strategies and specifies key variables that will guide future software developments and assist informatics decision-making in glycoproteomics.


Asunto(s)
Glicopéptidos/sangre , Glicoproteínas/sangre , Informática/métodos , Proteoma/análisis , Proteómica/métodos , Investigadores/estadística & datos numéricos , Programas Informáticos , Glicosilación , Humanos , Proteoma/metabolismo , Espectrometría de Masas en Tándem
8.
J Transl Med ; 22(1): 174, 2024 02 18.
Artículo en Inglés | MEDLINE | ID: mdl-38369502

RESUMEN

BACKGROUND: Overexpression of T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3) is related to the exhaustion of CD8+ tumor-infiltrating lymphocytes (TILs) in diffuse large B-cell lymphoma (DLBCL). However, the mechanism of TIM3-mediated CD8+TILs exhaustion in DLBCL remains poorly understood. Therefore, we aimed to clarify the potential pathway involved in TIM3-mediated CD8+TILs exhaustion and its significance in DLBCL. METHODS: The expression of TIM3 and its correlation with CD8+TILs exhaustion, the key ligand of TIM3, and the potential pathway of TIM3-mediated CD8+TILs exhaustion in DLBCL were analyzed using single-cell RNA sequencing and validated by RNA sequencing. The biological significance of TIM3-related pathway in DLBCL was investigated based on RNA sequencing, immunohistochemistry, and reverse transcription-quantitative polymerase chain reaction data. Finally, the possible regulatory mechanism of TIM3-related pathway in DLBCL was explored using single-cell RNA sequencing and RNA sequencing. RESULTS: Our results demonstrated that CD8+TILs, especially the terminally exhausted state, were the major clusters that expressed TIM3 in DLBCL. Galectin-9, mainly expressed in M2 macrophages, is the key ligand of TIM3 and can induce the exhaustion of CD8+TILs through TIM3/Galectin-9 pathway. Meanwhile, high TIM3/Galectin-9 enrichment is related to immunosuppressive tumor microenvironment, severe clinical manifestations, inferior prognosis, and poor response to CHOP-based chemotherapy, and can predict the clinical efficacy of immune checkpoint blockade therapy in DLBCL. Furthermore, the TIM3/Galectin-9 enrichment in DLBCL may be regulated by the IFN-γ signaling pathway. CONCLUSIONS: Our study highlights that TIM3/Galectin-9 pathway plays a crucial role in CD8+TILs exhaustion and the immune escape of DLBCL, which facilitates further functional studies and could provide a theoretical basis for the development of novel immunotherapy in DLBCL.


Asunto(s)
Linfocitos T CD8-positivos , Galectinas , Receptor 2 Celular del Virus de la Hepatitis A , Linfoma de Células B Grandes Difuso , Humanos , Receptor 2 Celular del Virus de la Hepatitis A/metabolismo , Ligandos , Linfocitos Infiltrantes de Tumor , Linfoma de Células B Grandes Difuso/patología , Microambiente Tumoral , Galectinas/metabolismo
9.
Cell Tissue Res ; 396(2): 177-195, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38366086

RESUMEN

Cannabidiol is a major component of cannabis but without known psychoactive properties. A wide range of properties have been attributed to it, such as anti-inflammatory, analgesic, anti-cancer, anti-seizure and anxiolytic. However, being a fairly new compound in its purified form, little is known about cannabidiol brain entry, especially during development. Sprague Dawley rats at four developmental ages: embryonic day E19, postnatal day P4 and P12 and non-pregnant adult females were administered intraperitoneal cannabidiol at 10 mg/kg with [3H] labelled cannabidiol. To investigate the extent of placental transfer, the drug was injected intravenously into E19 pregnant dams. Levels of [3H]-cannabidiol in blood plasma, cerebrospinal fluid and brain were estimated by liquid scintillation counting. Plasma protein binding of cannabidiol was identified by polyacrylamide gel electrophoresis and its bound and unbound fractions measured by ultrafiltration. Using available RNA-sequencing datasets of E19 rat brain, choroid plexus and placenta, as well as P5 and adult brain and choroid plexus, expression of 13 main cannabidiol receptors was analysed. Results showed that cannabidiol rapidly entered both the developing and adult brains. Entry into CSF was more limited. Its transfer across the placenta was substantially restricted as only about 50% of maternal blood plasma cannabidiol concentration was detected in fetal plasma. Albumin was the main, but not exclusive, cannabidiol binding protein at all ages. Several transcripts for cannabidiol receptors were expressed in age- and tissue-specific manner indicating that cannabidiol may have different functional effects in the fetal compared to adult brain.


Asunto(s)
Encéfalo , Cannabidiol , Ratas Sprague-Dawley , Animales , Cannabidiol/farmacología , Cannabidiol/sangre , Femenino , Encéfalo/metabolismo , Embarazo , Ratas , Feto/metabolismo , Placenta/metabolismo , Animales Recién Nacidos
10.
Opt Express ; 32(11): 20066-20079, 2024 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-38859124

RESUMEN

In the field of machine vision, depth segmentation plays a crucial role in dividing targets into different regions based on abrupt changes in depth. Phase-shifting depth segmentation is a technique that extracts singular points to form segmentation lines by leveraging the phase-shifting invariance of singular points in different wrapped phase maps. This makes it immune to color, texture, and camera exposure. However, current phase-shifting depth segmentation techniques face challenges in the precision of segmentation. To overcome this issue, this paper proposes a singular points extraction technique by constructing a more comprehensive threshold with the help of the minimum period of the phase map. Taking full advantage of the proposed technique, mean-value points and order singular points are accurately filtered out, and the integrity of segmentation lines in high-curvature regions can be guaranteed. During optimization processing, the precision of segmentation is improved by employing a low-cost morphology-based optimization model. Simulation results demonstrate the segmentation accuracy reaches up to 98.58% even in a noisy condition. Experimental results on different objects indicate that the proposed method exhibits good generalization and robustness.

11.
Exp Physiol ; 109(3): 427-444, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38059686

RESUMEN

Solute carriers (SLCs) regulate transfer of a wide range of molecules across cell membranes using facilitative or secondary active transport. In pregnancy, these transporters, expressed at the placental barrier, are important for delivery of nutrients to the fetus, whilst also limiting entry of potentially harmful substances, such as drugs. In the present study, RNA-sequencing analysis was used to investigate expression of SLCs in the fetal (embryonic day 19) rat brain, choroid plexus and placenta in untreated control animals and following maternal paracetamol treatment. In the treated group, paracetamol (15 mg/kg) was administered to dams twice daily for 5 days (from embryonic day 15 to 19). In untreated animals, overall expression of SLCs was highest in the placenta. In the paracetamol treatment group, expression of several SLCs was significantly different compared with control animals, with ion, amino acid, neurotransmitter and sugar transporters most affected. The number of SLC transcripts that changed significantly following treatment was the highest in the choroid plexus and lowest in the brain. All SLC transcripts that changed in the placenta following paracetamol treatment were downregulated. These results suggest that administration of paracetamol during pregnancy could potentially disrupt fetal nutrient homeostasis and affect brain development, resulting in major consequences for the neonate and extending into childhood.


Asunto(s)
Acetaminofén , Placenta , Humanos , Embarazo , Femenino , Animales , Ratas , Niño , Acetaminofén/farmacología , Plexo Coroideo , Feto , Encéfalo
12.
Langmuir ; 40(36): 19239-19250, 2024 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-39189153

RESUMEN

Supercapacitors with transition bimetallic oxides as pseudocapacitive materials have been of wide concern for their excellent energy storage performance. In this work, a simple coprecipitation method was used to synthesize the precursor, followed by calcination to prepare Ni-Cu bimetallic oxide materials. The structure, morphology and properties of the materials prepared by different precipitating agents and different calcination temperatures of NCO-H2C2O4 precursor were investigated. The optimum precipitant was determined to be H2C2O4, and Ni-Cu nanoparticles with regular lamellar microstructure were obtained at the calcination temperature of 400 °C. The nanostructure and morphology provide a large active channel for the rapid diffusion of electrolyte ions, and the specific capacitance of NCO-H2C2O4-400 electrode material can reach 740.31 F/g Cs at 1 A/g. The investigation of charge storage mechanism shows that the contribution rate of capacitance and diffusion control is about 37.9% and 67.2%, respectively. The electrochemical test results of the asymmetric supercapacitors (ASC) constructed with NCO-H2C2O4-400 and activated carbon show that the specific capacitance, energy density, and power density of the capacitor are 52.66 F/g, 16.45 Wh/kg, and 759.51 W/kg, respectively. Even after 5000 charge/discharge cycles at 5 A/g, it can still keep 90.57% of its initial capacity. This work not only provides competitive electrode materials for energy storage devices but also provides a feasible strategy for producing complex transition metal oxide materials with high capacitance performance.

13.
J Chem Phys ; 160(19)2024 May 21.
Artículo en Inglés | MEDLINE | ID: mdl-38757619

RESUMEN

The singlet fission process involves the conversion of one singlet excited state into two triplet states, which has significant potential for enhancing the energy utilization efficiency of solar cells. Carotenoid, a typical π conjugated chromophore, exhibits specific aggregate morphologies known to display singlet fission behavior. In this study, we investigate the singlet fission process in lycopene H-aggregates using femtosecond stimulated Raman spectroscopy aided by quantum chemical calculation. The experimental results reveal two reaction pathways that effectively relax the S2 (11Bu+) state populations in lycopene H-aggregates: a monomer-like singlet excited state relaxation pathway through S2 (11Bu+) → 11Bu- → S1 (21Ag-) and a dominant sequential singlet fission reaction pathway involving the S2 (11Bu+) state, followed by S* state, a triplet pair state [1(TT)], eventually leading to a long lifetime triplet state T1. Importantly, the presence of both anionic and cationic fingerprint Raman peaks in the S* state is indicative of a substantial charge-transfer character.

14.
Int J Mol Sci ; 25(16)2024 Aug 21.
Artículo en Inglés | MEDLINE | ID: mdl-39201766

RESUMEN

Sulfate transporters (SULTRs) are essential for the transport and absorption of sulfate in plants and serve as critical transport proteins within the sulfur metabolism pathway, significantly influencing plant growth, development, and stress adaptation. A bioinformatics analysis of SULTR genes in soybean was performed, resulting in the identification and classification of twenty-eight putative GmSULTRs into four distinct groups. In this study, the characteristics of the 28 GmSULTR genes, including those involved in collinearity, gene structure, protein motifs, cis-elements, tissue expression patterns, and the response to abiotic stress and plant hormone treatments, were systematically analyzed. This study focused on conducting a preliminary functional analysis of the GmSULTR3;1a gene, wherein a high expression level of GmSULTR3;1a in the roots, stems, and leaves was induced by a sulfur deficiency and GmSULTR3;1a improved the salt tolerance. A further functional characterization revealed that GmSULTR3;1a-overexpressing soybean hairy roots had higher SO42-, GSH, and methionine (Met) contents compared with the wild-type (WT) plant. These results demonstrate that the overexpression of GmSULTR3;1a may promote the sulfur assimilation metabolism and increase the content of sulfur-containing amino acids in plants.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Glycine max , Proteínas de Plantas , Estrés Fisiológico , Transportadores de Sulfato , Glycine max/genética , Glycine max/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estrés Fisiológico/genética , Transportadores de Sulfato/genética , Transportadores de Sulfato/metabolismo , Familia de Multigenes , Filogenia , Azufre/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/genética , Tolerancia a la Sal/genética , Sulfatos/metabolismo , Plantas Modificadas Genéticamente/genética , Perfilación de la Expresión Génica
15.
Angew Chem Int Ed Engl ; : e202408960, 2024 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-39212257

RESUMEN

Phosphonic acid (PA) self-assembled molecules have recently emerged as efficient hole-extraction layers (HELs) for organic solar cells (OSCs). However, the structural effects of PAs on their self-assembly behaviors on indium tin oxide (ITO) and thus photovoltaic performance remain obscure. Herein, we present a novel class of PAs, namely "non-fused ring dipodal phosphonic acids" (NFR-DPAs), featuring simple and malleable non-fused ring backbones and dipodal phosphonic acid anchoring groups. The efficacy of configurational isomerism in modulating the photoelectronic properties and switching molecular orientation of PAs atop electrodes results in distinct substrate surface energy and electronic characteristics. The NFR-DPA with linear (C2h symmetry) and brominated backbone exhibits favorable face-on orientation and enhanced work function modification capability compared to its angular (C2v symmetry) and non-brominated counterparts. This makes it versatile HELs in mitigating interfacial resistance for energy barrier-free hole collection, and affording optimal active layer morphology, which results in an impressive efficiency of 19.11% with a low voltage loss of 0.52 V for binary OSC devices and an excellent efficiency of 19.66% for ternary OSC devices. This study presents a new dimension to design PA-based HELs for high-performance OSCs.

16.
J Am Chem Soc ; 145(4): 2035-2039, 2023 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-36649589

RESUMEN

Experimental elucidation of the decoupling of electron and proton transfer at a molecular level is essential for thoroughly understanding the kinetics of heterogeneous (photo)electrochemical proton-coupled electron transfer water oxidation. Here we illustrate the electron-transfer intermediates of positively charged surface oxygenated species on Au (Au-OH+) and their correlations with the rate of water oxidation by in situ microphotoelectrochemical surface-enhanced Raman spectroscopy (SERS) and ambient-pressure X-ray photoelectron spectroscopy. At the intermediate stage of water oxidation, a characteristic blue shift of the vibration of Au-OH species in laser-power-density-dependent measurements was assigned to the light-induced production of Au-OH+ in water oxidation. The photothermal effect was excluded according to the vibrational frequencies of Au-OH species as the temperature was increased in a variable-temperature SERS measurement. Density functional theory calculations evidenced that the frequency blue shift is from the positively charged Au-OH species. The photocurrent-dependent frequency blue shift indicated that Au-OH+ is the key electron-transfer intermediate in water oxidation by decoupled electron and proton transfer.

17.
Anal Chem ; 95(50): 18388-18397, 2023 12 19.
Artículo en Inglés | MEDLINE | ID: mdl-38069741

RESUMEN

Determination of the relative expression levels of the α2,3/α2,6-sialic acid linkage isomers on glycoproteins is critical to the analysis of various human diseases such as cancer, inflammation, and viral infection. However, it remains a challenge to separate and differentiate site-specific linkage isomers at the glycopeptide level. Some derivatization methods on the carboxyl group of sialic acid have been developed to generate mass differences between linkage isomers. In this study, we utilized chemical derivatization that occurred on the vicinal diol of sialic acid to separate linkage isomers on a reverse-phase column using a relatively short time. 2-Aminobenzamide (2AB) labeling derivatization, including periodate oxidation and reductive amination, took only ∼3 h and achieved high labeling efficiency (>90%). Within a 66 min gradient, the sialic acid linkage isomers of 2AB-labeled glycopeptides from model glycoproteins can be efficiently resolved compared to native glycopeptides. Two different methods, neuraminidase digestion and higher-energy collision dissociation tandem mass spectrometry (HCD-MS2) fragmentation, were utilized to differentiate those isomeric peaks. By calculating the diagnostic oxonium ion ratio of Gal2ABNeuAc and 2ABNeuAc fragments, significant differences in chromatographic retention times and in mass spectral peak abundances were observed between linkage isomers. Their corresponding MS2 PCA plots also helped to elucidate the linkage information. This method was successfully applied to human blood serum. A total of 514 2AB-labeled glycopeptide structures, including 152 sets of isomers, were identified, proving the applicability of this method in linkage-specific structural characterization and relative quantification of sialic acid isomers.


Asunto(s)
Ácido N-Acetilneuramínico , Espectrometría de Masas en Tándem , Humanos , Ácido N-Acetilneuramínico/química , Espectrometría de Masas en Tándem/métodos , Sialoglicoproteínas , Cromatografía Líquida con Espectrometría de Masas , Cromatografía Liquida , Glicoproteínas , Glicopéptidos/análisis , Polisacáridos/química
18.
Anal Chem ; 95(44): 16059-16069, 2023 11 07.
Artículo en Inglés | MEDLINE | ID: mdl-37843510

RESUMEN

The complexity and heterogeneity of protein glycosylation present an analytical challenge to the studies of characterization and quantitation. Various LC-MS-based quantitation strategies have emerged in recent decades. Metabolic stable isotope labeling has been developed to enhance the accurate LC/MS-based quantitation between different cell lines. Stable isotope labeling by amino acids in a cell culture (SILAC) is the most widely used metabolic labeling method in proteomic analysis. However, it can only label the peptide backbone and is thus limited in glycomic studies. Here, we present a metabolic isotope labeling strategy, named GlyProSILC (Glycan Protein Stable Isotope Labeling in Cell Culture), that can label both the glycan motif and peptide backbone from the same batch of cells. It was performed by feeding cells with a heavy medium containing amide-15N-glutamine, 13C6-arginine (Arg6), and 13C6-15N2-lysine (Lys8). No significant change of cell line metabolism after GlyProSILC labeling was observed based on transcriptomic, glycomic, and proteomic data. The labeling conditions, labeling efficiency, and quantitation accuracy were investigated. After quantitation correction, we simultaneously quantified 62 N-glycans, 574 proteins, and 344 glycopeptides using the same batch of mixed 231BR/231 cell lines. So far, GlyProSILC provides an accurate and effective quantitation approach for glycomics, proteomics, and glycoproteomics in a cell culture system.


Asunto(s)
Glicómica , Proteómica , Marcaje Isotópico/métodos , Glicómica/métodos , Proteómica/métodos , Proteínas , Técnicas de Cultivo de Célula , Glicopéptidos/metabolismo , Polisacáridos/química
19.
Inorg Chem ; 62(19): 7212-7219, 2023 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-37139601

RESUMEN

Developing a photosensitizer with high efficiency and long-term stability for photocatalytic hydrogen evolution is highly desirable yet remains a challenge. Herein, a novel Ir(III) complex-based photosensitizer (Ir3) bearing coumarin and triphenylamine groups is designed. Ir3 exhibits record activity and durability among reported transition metal complexes for photocatalytic hydrogen evolution, with a TON of 198,363 and a duration of 214 h. The excellent photocatalytic performance of Ir3 can be attributed to the synergistic effect of coumarin and triphenylamine, which improves the visible light absorption, charge separation, and electron transfer capacity of photosensitizers. This is an efficient and long-lived Ir(III) photosensitizer constructed on the basis of a synergistic approach, which could provide a new insight for the development of high-performance Ir(III) photosensitizers at the molecular level.

20.
Macromol Rapid Commun ; 44(1): e2200297, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-35621302

RESUMEN

Molecules based on benzimidazolone-dioxazine are known as blue/violet pigments and have been commercialized for decades. However, unfavorable solubility limits the application of these structures as building blocks of conjugated polymers despite their low band gaps. Herein, a series of donor-acceptor conjugated polymers containing soluble benzimidazolone-dioxazine structures as the acceptors and oligothiophene as donors are synthesized and investigated. With increasing numbers of thiophene rings, the steric hindrance diminishes and high molecular weight polymers can be achieved, leading to an improved performance in organic field effect transistor devices. The hole mobility of polymers with three to six thiophene units is in the order of 10-1 cm2 V-1 s -1 . Among all the polymers, polymer P3 with three thiophene units between benzimidazolone-dioxazine structures shows the best hole mobility of 0.4 cm2 V-1 s -1 . Grazing-incidence wide-angle X-ray scattering results reveal that the high mobility of organic field-effect transistors (OFETs) can be accredited by matched donor-acceptor packing in the solid thin films.


Asunto(s)
Vendajes , Bencimidazoles , Polímeros , Tiofenos
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