Glucosaminyl Transferase Research Articles

Review on Molecular Cross-talk of Biofilm Producing Mechanisms of Staphylococcus aureus

Here the review converses the `molecular cross-talk` of biofuel production mechanisms for Staphylococcus aureus. Staphylococcus aureus is a leading cause of bacterial infections globally in both healthcare and community settings. The succes of this bacterium is the of an expansive repertoire of virulence factors in combination with acquired antibiotic resistance and propensity for biofilm formation. S. aureus leverages these factors to adapt to and subvert the host immune response. With the burgeoning fiels of immunometabolism, is has become clear that the metabolic program of leukocytes dictates their inflammatory status and overall effectiveness is clearing an infection. The treatment of S. aureus infections become complicated due to the capacity of S. aureus �multidrug-resistant� occurs because of biofilm formationon the surfaces depending on biotic and abiotic factors, genetic factors, and numerous environmental, which vary from species to species. A broad range of molecular phenomenon contributes a high range of recalcitrance that is insisting on the biofilm formation. The previous published literature illustrated that all strains of Staphylococcal sp. contain the �ica locus� and several can form biofilms in vitro condition. Absences of �ica locus� results diminish of capability to produce biofuels, along with `PIA gene`, or mediate `N-acetyl glucosaminyl transferase activity� in vitro condition.

APEX1 regulates alternative splicing of key tumorigenesis genes in non-small-cell lung cancer

BackgroundAberrant alternative splicing (AS) contributes to tumor progression. Previous studies have shown that apurinic-apyrimidinic endonuclease-1 (APEX1) is involved in tumor progression. It is unknown whether APEX1 functions in tumor progression by regulation of AS. It is also unknown whether APEX1 can regulate non-small-cell lung cancer (NSCLC) proliferation and apoptosis. We analyzed APEX1 expression levels in 517 lung NSCLC samples from the TCGA (Cancer Genome Atlas) database. The impact of APEX1 over expression on A549 cell proliferation and apoptosis was detected by the methyl thiazolyl tetrazolium assay and by flow cytometry. The transcriptome of A549 cells with and without APEX1 over expression was determined by Illumina sequencing, followed by analysis of AS. RT-qPCR validated expression of APEX1-related genes in A549 cells. We have successfully applied RNA-seq technology to demonstrate APEX1 regulation of AS.ResultsAPEX1 expression was shown to be upregulated in NSCLC samples and to reduce cell proliferation and induce apoptosis of A549 cells. In addition, APEX1 regulated AS of key tumorigenesis genes involved in cancer proliferation and apoptosis within MAPK and Wnt signaling pathways. Each of these pathways are involved in lung cancer progression. Furthermore, validated AS events regulated by APEX1 were in key tumorigenesis genes; AXIN1 (axis inhibition protein 1), GCNT2 (N-acetyl glucosaminyl transferase 2), and SMAD3 (SMAD Family Member 3). These genes encode signaling pathway transcription regulatory factors.ConclusionsWe found that increased expression of APEX1 was an independent prognostic factor related to NSCLC progression. Therefore, APEX1 regulation of AS may serve as a molecular marker or therapeutic target for NSCLC treatment.

Effects of 1,25(OH)2 D3 on lipid droplet growth in adipocytes.

This study aimed to explore the effects of 1,25(OH)2 D3 on lipid droplet (LD) growth in 3T3-L1 adipocytes of hypertrophy model. Cocktail method was used to induce differentiation in 3T3-L1 cells. After 8 days, the cells were modeled by 100, 300, 600, and 900 μM palmitic acid (PA) for 24 hr. The best concentration of modeling was screened by MTT results and triglycerides (TG) content. The model cells were intervened by 1, 10, and 100 nM 1,25(OH)2 D3 for 24 hr. Then, the TG content of cells were detected and stained by oil red O. The diameter and quantity of LDs were analyzed. mRNA relative expression levels of genes related to LD (CIDE-a, Fsp27, PLIN-1), upstream response factor (PPAR-α, PPAR-γ, and VDR), and TG metabolism (long chain acyl-CoA synthetase 3, 1-acylglycerol-3-phosphate O-acyltransferase 1, adipose triglyceride lipase, diacylglycerol acyltransferase 1, diacylglycerol acyltransferase 2, glycerol-3-phosphate O-acyltransferase 3, glycerol-3-phosphate O-acyltransferase 4, hormone-sensitive lipase, mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetyl glucosaminyl transferase, phosphatidic acid phosphatase, and uncoupling protein-1) were detected by RT-qPCR. A total of 300 μM PA was selected as the optimum concentration. Compared with model group, 10 and 100 nM 1,25(OH)2 D3 decreased the average diameter, increased the quantity of LDs, upregulated PPAR-α and PLIN-1 mRNA expression levels, and downregulated CIDE-a and Fsp27 mRNA expression levels significantly (p < .05). However, 1 nM 1,25(OH)2 D3 did not alter LD morphology and TG content. mRNA expression levels of long chain acyl-CoA synthetase 3, 1-acylglycerol-3-phosphate O-acyltransferase 1, diacylglycerol acyltransferase 2, glycerol-3-phosphate O-acyltransferase 3, and glycerol-3-phosphate O-acyltransferase 4 in 10 and 100 nM groups were significantly lower than those in the model group (p < .05); mRNA expression levels of adipose triglyceride lipase, diacylglycerol acyltransferase 1, hormone-sensitive lipase, mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetyl glucosaminyl transferase, phosphatidic acid phosphatase, and uncoupling protein-1 were significantly increased in the 100 nM group (p < .05). The 10 and 100 nM 1,25(OH)2 D3 can inhibit LD fusion, promote LD decomposition, reduce LD volume, and inhibit lipogenesis through the PPAR-α signaling pathway.

TGF-β1 Causes EMT by Regulating N-Acetyl Glucosaminyl Transferases via Downregulation of Non Muscle Myosin II-A through JNK/P38/PI3K Pathway in Lung Cancer.

Epithelial to mesenchymal transition (EMT) is a major determinant of cancer metastasis and is closely linked with TGF-β1. Intracellular proteins, including E. Cadherin, N. Cadherin and Vimentin are directly related to EMT that affect cell migration and adhesion; on the other hand, non muscle myosin (NM) has a central role in cytokinesis, migration and adhesion. We aimed to explore the association of EMT and metastasis with TGF-β1 through regulation of non-muscle myosin II-A (NMII-A) and its interaction with Hexosamine Biosynthesis Pathway (HBP). Protein expression changes were assessed by western blotting and immunofluorescent staining while transcription level changes were assessed by qRT-PCR. EMT was assessed by phenotypic analysis, wound healing, proliferation and transwell migration assay in vitro while in vivo studies were conducted in BALB/c nude mice for lung orthotopic and tail vein metastasis models. We demonstrated that regulation of JNK/ P38/PI3K by TGF-β1 led to down expression of NMII-A which promoted EMT and lung cancer metastasis. This down expression of NMII-A conversely upregulated the expression of Core 2 N-acetyl Glucosaminyl Transferase mucin type (C2GnT-M) and further facilitated up-regulation and down-regulation of N-acetylglucosaminyltransferase (GnT) -V and -III respectively; moreover, NMII-A K.D cells showed 3 times more tendency to migrate towards brain in vivo. The study reports a novel pathway through which NMII-A negatively regulates EMT and metastasis via up regulation of C2GnT-M, GnT-V and down expression of GnT-III. These findings of lung cancer may further be required to study other cancer types.

Prevention of IcaA regulated poly N-acetyl glucosamine formation in Staphylococcus aureus biofilm through new-drug like inhibitors: In silico approach and MD simulation study.

The effectiveness of various ligands against the protein structure of IcaA of the IcaABCD gene locus of Staphylococcus aureus were examined using the approach of structure based drug designing in reference with the protein's efficiency to form biofilms. Four compounds CID42738592, CID90468752, CID24277882, and CID6435208 were secluded from a database of 31,242 inhibitory ligands on the justification of the evaluated values falling under the four - tier structure based virtual screening. Under this principle value of least binding energy, human oral absorption and ADME properties were taken into consideration. Using the Glide module of Schrödinger, the above mentioned ligands showed an effective action against the protein IcaA which showed reduced activity as a glucosaminyl transferase. The complex of protein and ligand with best docking score was chosen for simulation studies. Structure based drug designing for the protein IcaA has given us potential leads as anti - biofilm agents. These screened out ligands might enable the development of new therapeutic strategies aimed at disrupting Staphylococcus aureus biofilms. The complex was showing stability towards the end of time for which it has been put for simulation. Thus molecule could be considered for making of biofilms.

Preparation and application of a ‘clickable’ acceptor for enzymatic synthesis of heparin oligosaccharides

A ‘clickable’ disaccharide was prepared by treating the aldehyde precursor with hydroxylamine, followed by the catalytic hydrogenation and diazotransfer reaction. This disaccharide was successfully applied to the elongation of the backbone construction of ultralow molecular weight (ULMW) heparins using two bacterial glycosyl transferases, N-acetyl glucosaminyl transferase from Escherichia coli K5 (KfiA) and heparosan synthase-2 (pmHS2) from Pasteurella multocida.

Uncovering Biphasic Catalytic Mode of C5-epimerase in Heparan Sulfate Biosynthesis

Heparan sulfate (HS), a highly sulfated polysaccharide, is biosynthesized through a pathway involving several enzymes. C(5)-epimerase (C(5)-epi) is a key enzyme in this pathway. C(5)-epi is known for being a two-way catalytic enzyme, displaying a "reversible" catalytic mode by converting a glucuronic acid to an iduronic acid residue, and vice versa. Here, we discovered that C(5)-epi can also serve as a one-way catalyst to convert a glucuronic acid to an iduronic acid residue, displaying an "irreversible" catalytic mode. Our data indicated that the reversible or irreversible catalytic mode strictly depends on the saccharide substrate structures. The biphasic mode of C(5)-epi offers a novel mechanism to regulate the biosynthesis of HS with the desired biological functions.

Isolation of a Mutant Arabidopsis Plant That Lacks N-Acetyl Glucosaminyl Transferase I and Is Unable to Synthesize Golgi-Modified Complex N-Linked Glycans

The complex asparagine-linked glycans of plant glycoproteins, characterized by the presence of beta 1-->2 xylose and alpha 1-->3 fucose residues, are derived from typical mannose9(N-acetylglucosamine)2 (Man9GlcNAc2) N-linked glycans through the activity of a series of glycosidases and glycosyl transferases in the Golgi apparatus. By screening leaf extracts with an antiserum against complex glycans, we isolated a mutant of Arabidopsis thaliana that is blocked in the conversion of high-manne to complex glycans. In callus tissues derived from the mutant plants, all glycans bind to concanavalin A. These glycans can be released by treatment with endoglycosidase H, and the majority has the same size as Man5GlcNAc1 glycans. In the presence of deoxymannojirimycin, an inhibitor of mannosidase I, the mutant cells synthesize Man9GlcNAc2 and Man8GlcNAc2 glycans, suggesting that the biochemical lesion in the mutant is not in the biosynthesis of high-mannose glycans in the endoplasmic reticulum but in their modification in the Golgi. Direct enzyme assays of cell extracts show that the mutant cells lack N-acetyl glucosaminyl transferase I, the first enzyme in the pathway of complex glycan biosynthesis. The mutant plants are able to complete their development normally under several environmental conditions, suggesting that complex glycans are not essential for normal developmental processes. By crossing the complex-glycan-deficient strain of A. thaliana with a transgenic strain that expresses the glycoprotein phytohemagglutinin, we obtained a unique strain that synthesizes phytohemagglutinin with two high-mannose glycans, instead of one high-mannose and one complex glycan.

Effectivity of dolichyl phosphates with different chain lengths as acceptors of nucleotide activated sugars

Chemical synthesis of different S-forms of dolichyl-P was performed in order to investigate the use of these polyprenes in mannosyl, glucosyl and glucosaminyl transferase reactions. Determination of the Vmax values for a series of dolichyl-P demonstrated that the velocities of transferase reactions with all those dolichyl-P derivatives present in animal tissues are largely the same. The apparent Km values for the various dolichyl-P in the transferase system studied differed, but this property does not appear to have physiological importance.

Stimulation of UDP-N acetyl glucosamine: Glycoprotein N-acetyl glucosaminyl transferase activity by cytidine 5′-diphospho-choline

Golgi-rich and golgi-depleted heterogenous membrane fractions were prepared from livers of rats fed choline supplemented or deficient diets. Assay of UDP-N-acetyl glucosamine: glycoprotein N-acetyl glucosaminyl transferase activity showed a decrease of the enzyme activity in the golgi-depleted membrane fraction of the choline deficient liver. A marked stimulation of the enzyme activity in the same fraction was observed by in vitro additions of phosphorylcholine plus cytidine 5′-triphosphate or cytidine 5′-diphospho-choline. This stimulation was found to be dose dependent. Two possible mechanisms are suggested for the observed effect.