SEARCH · PubMed Health

Results for “GLUCOSYLTRANSFERASES”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,045 records · Page 58Linked to original sources

PubMed1982

Changes in collagen metabolism in diseased muscle. I. Biochemical studies.

Possible changes in collagen biosynthesis were studied in 50 patients with neuromuscular disorders and 14 controls. Type III procollagen aminoterminal propeptide concentrations and galactosylhydroxylysyl glucosyltransferase (GGT) activities were assayed in serum, and prolyl 4-hydroxylase and GGT activities were assayed in muscle biopsy specimens. All four assays showed significantly elevated values in cases of polymyositis, adult forms of muscular dystrophy, and amyotrophic lateral sclerosis, the concentration of muscular collagen also being significantly increased in the last two conditions. Some abnormalities were also seen in polyneuropathy, myotonia congenita, and undefined myopathy. High correlations were found among the values for the four assays, but no marked correlations with muscular collagen concentration or enzyme activities characteristic of neuromuscular disorders were found. The four assays may reflect changes in actual collagen synthesis in the diseased muscle.

Adult↗

PubMed2000

The effect of 50 Hz magnetic field on GCSmRNA expression in lymphoma B cell by mRNA differential display.

Magnetic fields (MFs) of various characteristics can lead to plethora effects in biological system. From a molecular point of view, we hypothesized that there must be a fundamental difference in gene expression between the MF exposed and the unexposed cell. To identify the classes of genes that are regulated, 0.8 mT 50 Hz MF-induced changes in gene expression were examined in a Daudi cell culture using differential display and reverse transcriptase-polymerase chain reaction. A candidate cDNA (signatured as MF-CB) that was observed in the sham-exposed but not in MF-exposed cultures was recovered and reamplified. After verification by Northern blot, the cDNA was cloned and sequenced. It was found that 254-base pair of 5'-end MF-CB cDNA clone was identical to gcs in open reading frame (ORF) range. Based on the preliminarily sequence, the prolonged length of 5'-end MF-CB cDNA was obtained by PCR amplification and its sequence analysis showed the same results as its original fragment. In order to further determine whether MF-CB cDNA is from gcs, two Northern blots were probed with gcs and MF-CB cDNA, respectively, and the data revealed signals of the same size and expression pattern on the two probe filters, which demonstrated that MF-CB is an EST (expression sequence tag) of gcs. gcs is a gene, identified recently (GenBank accession number D89866), encoding ceramide glucosyltransferase (GCS), which has been implicated as a causal element in human cell growth and differentiation. In an additional experiment, time-dependent changes in the transcription of gcs induced by 0.8 mT MF were observed by Northern blot with a sharp and reproducible inhibition effect after 20 min exposure and a reduction after 20-24 h exposure. The study demonstrates for the first time that 50 Hz MF can lead to changes in gcs transcription, which provides a new clue to elucidate the mechanism by which MF influence cell growth and differentiation.

Amino Acid Sequence↗

PubMed2000

Effect of glucosphingolipid synthesis inhibitor (PPMP and PDMP) treatment on Tetrahymena pyriformis: data on the evolution of the signaling system.

1-Phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) and 1-phenyl-2-palmitoylamino-3-morpholino-1-propanol (PPMP) are structural analogues of ceramide; inhibiting UDP-glucose : ceramide glucosyltransferase. After treatment with these synthetic ceramide analogues the expression of glucosphingolipid decreases, while ceramide and sphingomyelin levels increase in the cells of higher eukaryotes. In the unicellular Tetrahymena pyriformis, treatment with PDMP (10-20 microM) and PPMP (40-80 microM) influenced the synthesis of galactose, glucosamine and mannose-containing lipids. On the whole the amount of these lipids was reduced, but new galactose and glucosamine-containing lipids appeared (the exact structures of these lipids were not characterized). Incorporation of (32)P into phosphatidylethanolamine (PE) and phosphatidic acid (PA) was decreased significantly; however the amount of inositol phospholipids were increased. The incorporation of 3H-serine into phosphatidylserine was abolished, but incorporation into sphingomyelin and ceramide was increased. The cytoskeletal elements (silver line system) were disturbed on the basis of scanning electron microscopic pictures. The TRITC-Con A binding and the morphology of the cells were influenced as revealed by confocal laser scanning microscopic analyses. In contrast to higher eukaryotes, in Tetrahymena the shorter fatty acyl chain variant (PDMP) proved to be more effective in each of the examined parameters, while the longer chain variants (PPMP) had milder activity.

Animals↗

PubMed2005

Clinical and molecular characterization of the first adult congenital disorder of glycosylation (CDG) type Ic patient.

Congenital disorder of glycosylation (CDG) type Ic, the second largest subtype of CDG, is caused by mutations in human ALG6 (hALG6). This gene encodes the alpha1,3-glucosyltransferase that catalyzes transfer of the first glucose residue to the lipid-linked oligosaccharide precursor for N-linked glycosylation. In this report, we describe the first adult patient diagnosed with CDG-Ic, carrying two previously unknown mutations. The first is a three base deletion (897-899delAAT) leading to the loss of I299, the second is an intronic mutation (IVS7 + 2T > G) that causes aberrant splicing. Wildtype hALG6, delivered by a lentiviral vector into patient's fibroblasts, clearly improves the biochemical phenotype, which confirms that the mutations are disease-causing. Striking clinical findings include limb deficiencies in the fingers, resembling brachydactyly type B, a deep vein thrombosis, pseudotumor cerebri, and endocrine disturbances with pronounced hyperandrogenism and virilization. However, even in adulthood, this patient shows normal magnetic resonance imaging of the brain.

Adult↗

PubMed2006

Effect of partially hydrolyzed soluble glucan produced by glucosyltrasferases of Streptococcus mutans on stimulating human T cell.

Soluble glucan, which was obtained from action of glucosyltransferases (GTFs) of Streptococcus mutans on sucrose, was partially hydrolyzed by acetic acid and examined for human T lymphoblast (MOLT-4) stimulating activity. Addition of the partially hydrolyzed glucan (15-60 microg/ml) stimulated human T cell (39-65%) in a dose dependant manner according to MTT assay. Production of interleukine-2 (IL-2) and interleukine-2 receptor (IL-2R) from T cell was increased by 44.5 and 25%, respectively, by addition of partially hydrolyzed glucan (15 microg/ml). These results indicate that stimulation of human T cells by hydrolyzed glucan is probably caused by its effects on stimulating gene expression of IL-2 and IL-2R of human T cell.

Acetic Acid↗

PubMed2001

Utilizing genetically engineered bacteria to produce plant-specific glucosides.

Plant-derived glucosides have attracted much attention due to their widespread applications. This class of products is difficult to isolate or to synthesize in pure form because of the resulting low yields. Thus, simple approaches for the generation of such glucosides would be highly beneficial. We purified and characterized a novel glucosyltransferase from plant cell suspension cultures of Rauvolfia serpentina, which showed rather low substrate specificity. We obtained its cDNA and expressed the active recombinant protein in bacteria (Escherichia coli) with excellent plant-specific glucosylation efficiencies. Compared with the plant system, the bacteria delivered the new enzyme, which was in the form of a soluble or matrix-bound enzyme, approximately 1800 times more efficiently for the synthesis of a wide range of glucosides. More importantly, the engineered E. coli strain allowed for in vivo glucosylation and release of the product into the culture medium, as shown by the formation of arbutin, which is a potent inhibitor of human melanin biosynthesis with commercial value.

Arbutin↗

PubMed1979

Dextran biosynthesis and dextransucrase production by continuous culture of Leuconostoc mesenteroides.

Leuconostoc mesenteroides NRRL B-512(F) was grown in continuous culture under conditions of energy-limited growth. The extracellular enzyme dextransucrase (sucrose: 1,6-alpha-D-glucan 6-alpha-glucosyltransferase EC 2.4.1.5), was not detected in glucose- or maltose-limited cultures. Under conditions of sucrose-limited growth, the enzyme activity of the cell-free culture supernatant increased with increasing dilution rate only after the critical concentration of enzyme inducer (sucrose) in the chemostat had been achieved. The appearance of fructose in the effluent of the sucrose-limited chemostat at higher dilution rates indicated that sucrose was being diverted to dextran biosynthesis. The competition between bacteria and extracellular enzyme for the common substrate sucrose represents an inefficiency in the system of enzyme production. Dextransucrase was isolated from the cell-free culture supernatant by ammonium sulfate precipitation and DEAE-cellulose chromatography. The enzyme preparation exhibited both dextran biosynthetic activity and an invertase-like activity. The biosynthetic efficiency was increased by decreasing the temperature from 30 to 10 degrees C. The enzyme was irreversibly denatured by prolonged incubation in the absence of Ca2+.

Dextrans↗

PubMed2006

RebG- and RebM-catalyzed indolocarbazole diversification.

Rebeccamycin and staurosporine represent two broad classes of indolocarbazole glycoside natural products with antitumor properties. Based upon previous sequence annotation and in vivo studies, rebG encodes for the rebeccamycin N-glucosyltransferase, and rebM for the requisite 4'-O-methyltransferase. In the current study, an efficient in vivo biotransformation system for RebG was established in both Streptomyces lividans and Escherichia coli. Bioconversion experiments revealed RebG to glucosylate a set of indolocarbazole surrogates, the products of which could be further modified by in vitro RebM-catalyzed 4'-O-methylation. Both RebG and RebM displayed substrate promiscuity, and evidence for a remarkable lack of RebG regioselectivity in the presence of asymmetric substrates is also provided. In the context of the created indolocarbazole analogues, cytotoxicity assays also highlight the importance of 4'-O-methylation for their biological activity.

Bacterial Proteins↗

PubMed1989

Tissue-specific effects of maize bronze gene promoter mutations induced by Ds1 insertion and excision.

Bz-wm is an allele of the Bz locus of maize isolated by McClintock (1962) as a derivative of bz-m2. It contains a Ds1 insertion 63 bp upstream of the start of transcription and a 3 bp insertion in the coding region at the site of the Ac element that was present in bz-m2. Bz-wm produces, in the aleurone layer of the endosperm, low amounts (approximately 1% of wild-type) of a Bz-gene encoded UDP-glucose: flavoid 3-0-glucosyltransferase (UFGT) polypeptide with altered thermal stability. Three phenotypically wild-type derivatives, Bz' (wm)-1, Bz' (wm)-2 and Bz' (wm)-3, were isolated in the presence of Ac and shown to have excised the Ds1 element but not fully restored UFGT activity in endosperm assays. In the studies reported here, we have further analyzed these Bz' derivatives of Bz-wm by determining the DNA sequences left behind on Ds1 excision, and by measuring the amount of UFGT activity and/or Bz mRNA conditioned by Bz-wm and the Bz' derivatives in different tissues. The data indicate that tissue-specific differences in expression of the Bz gene have been produced in alleles with mutations caused by transposable elements Ac and Ds. These mutations may affect either the amount of Bz transcription or the stability of the UFGT polypeptide. The sequence or spacing in the -63 region of the Bz promoter appears to be critical for maximum expression in aleurone and husk but not in pollen and pigmented seedling tissue.

Alleles↗

PubMed1982

Enzymes of collagen synthesis and type III procollagen amino-propeptide in serum from Nigerians with hepato-cellular carcinoma and other malignant diseases.

Serum immunoreactive prolyl hydroxylase protein (S-IRPH), galactosylhydroxylysyl glucosyltransferase activity (S-GGT) and the amino-terminal propeptide of type III procollagen [S-Pro(III)-N-P] were studied in 24 patients with primary hepatocellular carcinoma, 18 with secondary liver neoplasms and 35 with other malignant diseases but no evidence of liver involvement; this latter group included 13 patients with Burkitt's lymphoma, 11 with breast cancer and 11 with other neoplasms. Control values were determined for 60 apparently healthy Nigerians, S-IRPH and S-GGT were above the upper normal limit, defined as the mean + 2 SD of the controls, in all the patients with primary hepatocellular carcinoma and all but one with secondary liver neoplasms, whereas only one S-IRPH value and three S-GGT values exceeded this limit in the patients with other malignant diseases. The mean S-Pro(III)-N-P was even more elevated than S-IRPH and S-GGT in the primary and secondary liver neoplasm cases, but was also elevated in other malignant neoplasms; about one third of the patients with no evidence of liver involvement had a concentration exceeding the upper normal limit. A high correlation was found between the values for the three assays both in primary hepatocellular carcinoma and in the whole series of malignant diseases. The data suggest that primary and secondary malignant neoplasms of the liver have a high rate of collagen synthesis. The three assays may be of some value in the diagnosis of primary hepatocellular carcinoma and secondary liver involvement in other malignant diseases, and in monitoring the treatment provided.

Adolescent↗

PubMed2006

Lysyl hydroxylase 3 (LH3) modifies proteins in the extracellular space, a novel mechanism for matrix remodeling.

Lysyl hydroxylase 3 (LH3), the multifunctional enzyme associated with collagen biosynthesis that possesses lysyl hydroxylase and collagen glycosyltransferase activities, has been characterized in the extracellular space in this study. Lysine modifications are known to occur in the endoplasmic reticulum (ER) prior to collagen triple-helix formation, but in this study we show that LH3 is also present and active in the extracellular space. Studies with in vitro cultured cells indicate that LH3, in addition to being an ER resident, is secreted from the cells and is found both in the medium and on the cell surface associated with collagens or other proteins with collagenous sequences. Furthermore, in vivo, LH3 is present in serum. LH3 protein levels correlate with the galactosylhydroxylysine glucosyltransferase (GGT) activity of mouse tissues. This, together with other data, indicates that LH3 is responsible for GGT activity in the tissues and that GGT activity assays can be used to quantify LH3 in tissues. LH3 in vivo is located in two compartments, in the ER and in the extracellular space, and the partitioning varies with tissue type. In mouse kidney the enzyme is located mainly intracellularly, whereas in mouse liver it is located solely in the extracellular space. The extracellular localization and the ability of LH3 to modify lysyl residues of extracellular proteins in their native, nondenaturated conformation reveals a new dynamic in extracellular matrix remodeling, suggesting a novel mechanism for adjusting the amount of hydroxylysine and hydroxylysine-linked carbohydrates in collagenous proteins.

Animals↗

PubMed1997

Effect of certain nutrients on the production of dextransucrase from Leuconostoc mesenteroides NRRL B-512F.

The effects of certain nutrients on dextransucrase (sucrose: 1,6-alpha-d-glucan 6-alpha-d-glucosyltransferase EC 2.4.1.5) production from Leuconostoc mesenteroides NRRL B-512F were studied. An increase in concentration of sucrose to 4% in the enzyme production medium resulted in the increase of activity of dextransucrase. Higher enzyme yields were obtained at low yeast extract and high phosphate concentrations. The presence of peptone and beef extract in the medium in addition to 2% yeast extract resulted in an enhanced production of dextransucrase. The enzyme activity increased by 30% by both peptone and beef extract. Addition of Tween 80 to the medium enhanced the production of enzyme and the activity was increased by 25%. Magnesium ions stimulated the activity marginally. Sodium fluoride enhanced the activity of dextransucrase by 25%.

Culture Media↗

PubMed1994

Collagens in the injured porcine intervertebral disc.

Spinal pain often is thought to be due to degeneration and mechanical failure of the intervertebral disc. Since the mechanical strength of the tissue depends on collagen fibers, the present study was designed to investigate the reactions in collagen metabolism after an experimentally induced disc injury. Five domestic pigs underwent an incision in the anterior part of the annulus fibrosus of disc L4-L5 through a retroperitoneal approach. The animals were killed 3 months postoperatively, and the injured discs and intact discs (controls) from different animals were removed for chemical analysis. Slices were cut from seven different parts across the disc. The concentration of total collagen (hydroxyproline [Hyp]), the activities of the two key enzymes in collagen biosynthesis (prolyl 4-hydroxylase [PH] and galactosylhydroxylysyl glucosyltransferase [GGT]), and the concentration of mature collagen crosslinks (hydroxypyridinium [HP]) were determined. In all experimental discs, the morphology had changed considerably: the nucleus pulposus was small, fibrous, and yellowish. The annular lamellar structure was partially destroyed and had been replaced by granulation tissue in the region of the injury. Large osteophytes had formed at the ventral edges of the vertebral bodies. In the nucleus pulposus, the Hyp concentration and the activities of PH and GGT were significantly increased, whereas the water content had decreased. The concentration of HP crosslinks was decreased in the anterior annulus fibrosus.

Animals↗

PubMed2002

Functional, comparative and cell biological analysis of Saccharomyces cerevisiae Kre5p.

Saccharomyces cerevisiae kre5delta mutants lack beta-1,6-glucan, a polymer required for proper cell wall assembly and architecture. A functional and cell biological analysis of Kre5p was conducted to further elucidate the role of this diverged protein glucosyltransferase-like protein in beta-1,6-glucan synthesis. Kre5p was found to be a primarily soluble N-glycoprotein of approximately 200 kDa, that localizes to the endoplasmic reticulum. The terminal phenotype of Kre5p-deficient cells was observed, and revealed a severe cell wall morphological defect. KRE6, encoding a glucanase-like protein, was identified as a multicopy suppressor of a temperature-sensitive kre5 allele, suggesting that these proteins may participate in a common beta-1,6-biosynthetic pathway. An analysis of truncated versions of Kre5p indicated that all major regions of the protein are required for viability. Finally, Candida albicans KRE5 was shown to partially restore growth to S. cerevisiae kre5delta cells, suggesting that these proteins are functionally related.

Alleles↗

PubMed1994

Mammalian glycosyltransferases prefer glycosyl phosphoryl dolichols rather than glycosyl phosphoryl polyprenols as substrates for oligosaccharyl synthesis.

We have studied the effectiveness of polyprenyl-P-mannose and polyprenol-P-glucose as donor substrates for the dolichyl-P-mannose:Man5(GlcNAc)2-PP-dolichol mannosyltransferase and the dolichyl-P-glucose:Man9(GlcNAc)2-PP-dolichol glucosyltransferase, respectively. The polyprenol moiety differs from dolichol only in the unsaturation of the terminal isoprene unit of the molecule. Based on the kinetics of the reactions, we have found that both glycosyltransferases have higher apparent Kms and lower apparent Vmaxs using polyprenyl-P-monosaccharides as substrates rather than the dolichyl-P-monosaccharides. The products formed with the polyprenyl-P-sugars were the same as those formed by the dolichol-linked sugars, indicating that the polyprenol substrates could be utilized by the glycosyltransferases in vitro. The results also indicate that the dolichyl-P-sugars and the polyprenyl-P-sugars compete for the same binding site on the enzyme. These findings are significant in terms of understanding the glycosylation phenotypes of Chinese hamster ovary cell mutants of the Lec9 complementation group, which lack the ability to convert polyprenol into dolichol.

Animals↗

PubMed1997

Ceramide glucosylation in bean hypocotyl microsomes: evidence that steryl glucoside serves as glucose donor.

The formation of glucosylceramide, the predominant sphingolipid in plant tissues, was examined in microsomes from wax bean hypocotyls. Membranes were incubated with UDP-[14C]glucose in an assay mixture. The lipid extracts obtained from the assays were separated by thin-layer chromatography, and the radioactivity incorporated into glucosylceramide, steryl glucoside, and acylated steryl glucoside was determined. Although the formation of glucosylceramide was detected and characterized, several lines of evidence contradicted the assumption that UDP-glucose is the immediate glucose donor for glucosylceramide formation in plants: PDMP (DL-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol), an inhibitor of ceramide glucosyltransferase in animal tissues, did not inhibit glucosylceramide formation in bean microsomes. Addition of UDP-glucose pyrophosphorylase during the assay to degrade UDP-[14C]glucose blocked the further production of labeled steryl glucoside, but did not prevent the continued formation of labeled glucosylceramide. Omitting UDP-[14C]glucose and including steryl [14C]glucoside in the assay resulted in the formation of labeled glucosylceramide. Collectively, these results suggest that glucosylceramide formation in plants does not utilize UDP-glucose as the immediate glucose donor, as has been demonstrated for the reaction in animal tissues, and that steryl glucoside serves as glucose donor for ceramide formation. This study, the first to examine glucosylceramide formation in plants, provides evidence for a novel enzymatic reaction in sphingolipid synthesis as well as a new, metabolic role for steryl glucoside in plant tissues.

Ceramides↗

PubMed1997

A microtiter-based fluorescence assay for (1,3)-beta-glucan synthases.

A high-throughput assay for UDP-Glc:(1,3)-beta-glucan synthase(EC 2.4.1.34, UDP-glucose:1,3-beta-D-glucan, 3-beta-glucosyltransferase) from fungi and higher plants is described. The assay is performed in microtiter plates and is extremely inexpensive compared to other standard assays for these enzymes. The reduction in price is achieved by replacing the conventional substrate UDP-[14C]Glc with its nonradioactive counterpart, and the nonradioactive glucan produced is quantified as a fluorescent complex following specific interaction with the fluorochrome present in commercial aniline blue. In addition to a > 100-fold reduction in cost, the assay is highly reproducible and nearly as sensitive as radioactive assays and has the additional advantages of increased safety and avoidance of the need for filtration and washing steps to collect the glucan product. As such, the assay is highly suitable for high-throughput screening for inhibitors of these enzymes.

Aniline Compounds↗

PubMed1996

Secretion of green fluorescent protein from recombinant baculovirus-infected insect cells.

Trichoplusia ni (High Five) and Spodoptera frugiperda (Sf21) cells were engineered for expression of epitope (Flag)-tagged signal peptide-green fluorescent protein (GFP) fusions to examine the suitability of GFP as a secretory marker. The recombinant baculovirus-infected cells became fluorescent, and the High Five cells but not Sf21 cells secreted GFP in the culture medium as detected by the presence in the culture supernatant of a Flag-immunoreactive 30-kDa species and the characteristic 510-nm GFP fluorescence peak. Signal peptides derived from ecdysteroid UDP-glucosyltransferase of Autographa californica nuclear polyhedrosis virus and from rat brain glutamate receptor were both able to promote secretion of GFP. GFP may thus be used as a research tool in the study of the secretory process in insect cells both in cell biology and in biotechnological applications.

Amino Acid Sequence↗