Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.15 (pectinase)
 2,440 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Studies were carried out to determine the distribution of the following: (1) carbamoyl phosphate synthetase (EC 2.7.2.9), (2) ornithine carbamoyltransferase (EC 2.1.3.3), (3) argininosuccinate synthetase (EC 6.3.4.5), and (4) argininosuccinate lyase (EC 4.3.2.1) in soybean cells grown in suspension culture. Protoplasts were produced from the soybean cells by treatment with cellulase (EC 3.2.1.4) and pectinase (EC 3.2.1.15); the protoplasts were then ruptured by osmotic shock with distilled water. This treatment was followed by differential centrifugation and sucrose density gradient centrifugation to isolate various organelle fractions including mitochondria and plastids. Examination of these fractions using specific enzyme assays showed that carbamoylphosphate synthetase and ornithine carbamoyltransferase were localized in a fraction found to be composed primarily of plastids. Argininosuccinate synthetase and argininosuccinate lyase appeared to be associated with either the cytosol or a membrane fraction in close association with the cytosol such as the endoplasmic reticulum or protoplast membrane.
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PMID:The localization within plant cells of enzymes involved in arginine biosynthesis. 56 67

The pyroantimonate method was used to study the localization of free and weakly bound calcium in cells of moss protonema of Funaria hygrometrica Hedw. cultivated on a clinostat (2rev/min). Electroncytochemical study of control cells cultivated at 1 g revealed that granular precipitate marked chloroplasts, mitochondria, Golgi apparatus, lipid drops, nucleoplasma, nucleolus, nucleus membranes, cell walls and endoplasmic reticulum. In mitochondria the precipitate was revealed in stroma, in chloroplast it was found on thylakoids and envelope membranes. The cultivation of protonema on clinostat led to the intensification in cytochemical reaction product deposit. A considerable intensification of the reaction was noted in endomembranes, vacuoles, periplasmic space and cell walls. At the same time analysis of pectinase localization was made using the electroncytochemical method. A high reaction intensity in walls in comparison to that in control was found out to be a distinctive peculiarity of the cells cultivated on clinostat. It testifies to the fact that increasing of free calcium concentrations under conditions of clinostation is connected with pectinic substances hydrolysis and breaking of methoxy groups of pectins. Data obtained are discussed in relation to problems of possible mechanisms of disturbance in calcium balance of plant cells and the role of cell walls in gomeostasis of cell grown under conditions of simulated weightlessness.
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PMID:Long clinostation influence on the localization of free and weakly bound calcium in cell walls of Funaria hygrometrica moss protonema cells. 1153 54

Alpha-1,4-galacturonosyltransferase (GalAT) is an enzyme required for the biosynthesis of the plant cell wall pectic polysaccharide homogalacturonan (HGA). GalAT activity in homogenates from pea (Pisum sativum L. var. Alaska) stem internodes co-localized in linear and discontinuous sucrose gradients with latent UDPase activity, an enzyme marker specific for Golgi membranes. GalAT activity was separated from antimycin A-insensitive NADH:cytochrome c reductase and cytochrome c oxidase activities, enzyme markers for the endoplasmic reticulum and the mitochondria, respectively. GalAT and latent UDPase activities were separated from the majority (80%) of callose synthase activity, a marker for the plasma membrane, suggesting that little or no GalAT is present in the plasma membrane. GalAT activities in proteinase K-treated and untreated Golgi vesicles were similar, whereas no GalAT activity was detected after treating Golgi vesicles with proteinase K in the presence of Triton X-100. These results demonstrate that the catalytic site of GalAT resides within the lumen of the Golgi. The products generated by Golgi-localized GalAT were converted by endopolygalacturonase treatment to mono- and di-galacturonic acid, thereby showing that GalAT synthesizes 1-->4-linked alpha-D-galacturonan. Our data provide the first enzymatic evidence that a glycosyltransferase involved in HGA synthesis is present in the Golgi apparatus. Together with prior results of in vivo labeling and immunocytochemical studies, these results show that pectin biosynthesis occurs in the Golgi. A model for the biosynthesis of the pectic polysaccharide HGA is proposed.
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PMID:The catalytic site of the pectin biosynthetic enzyme alpha-1,4-galacturonosyltransferase is located in the lumen of the Golgi. 1155 63

In order to facilitate production and secretion of a soluble form of a small, single-chain antibody ScFv (32 kDa) in tobacco cell suspension culture, several modifications were made simultaneously to the antibody cDNA that included elements that have been shown to regulate the expression of proteins in plants. The scFv cDNA was initially ligated into a binary vector under the control of the CaMV 35S promoter and the T7 terminator for expression in tobacco suspension culture. Subsequently, modifications were engineered into the cDNA for enhancement of scFv production. These included the following: (i) the signal peptide (SP) of the tobacco pathogenesis-related protein PR1a which was added in-frame to the N-terminal end of scFv cDNA; (ii) a 5'-nontranslated region from the tobacco etch virus (TEV leader sequence), which was fused to the N-terminal end of the SP; and (iii) the endoplasmic reticulum retention signal peptide KDEL, which was added to the C-terminal end of the scFv protein. Using a modified disruption method involving pectinase, the highest expression of total scFv (344 ng scFv/g cell) occurred when the plant leader sequence, the TEV sequence, and the KDEL peptide were all present in the expression construct. Although the addition of the KDEL sequence significantly increased the total yield of protein 5.4-fold, it did not increase the overall amount of protein secreted. These studies indicate that while the SP is very important in promoting secretion of the scFv, it had little influence on increasing scFv secretion levels even when both the TEV and the KDEL sequences significantly increased overall protein levels.
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PMID:Combined use of regulatory elements within the cDNA to increase the production of a soluble mouse single-chain antibody, scFv, from tobacco cell suspension cultures. 1192 54

Ultrastructural changes in the pericarp of tomato (Lycopersicon esculentum Mill) fruit were followed during ripening. Ethylene production was monitored by gas chromatography and samples analyzed at successive stages of the ripening process.Changes in the cytoplasmic ultrastructure were not consistent with the suggestion that ripening is a ;senescence' phenomenon. A large degree of ultrastructural organization, especially of the mitochondria, chromoplasts, and rough endoplasmic reticulum, was retained by ripe fruit.Striking changes in the structure of the cell wall were noted, beginning with dissolution of the middle lamella and eventual disruption of the primary cell wall. These changes were correlated with appearance of polygalacturonase (EC 3.2.1.15) isoenzymes. Application of purified tomato polygalacturonase isoenzymes to mature green fruit tissue duplicated the changes in the cell wall noted during normal ripening. Possible roles of the polygalacturonase isoenzymes in cell wall disorganization are discussed.
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PMID:Ultrastructure of tomato fruit ripening and the role of polygalacturonase isoenzymes in cell wall degradation. 1666 25

It is widely acknowledged that plant-made pharmaceuticals (PMPs) offer numerous benefits, including inexpensive production, biological safety and the facility for production at agricultural scale. At the same time, it is important to minimize any potential risk associated with this new technology, including the potential release of bioactive proteins into the environment. To address this issue, we studied transgenic Nicotiana benthamiana and Nicotiana tabacum plants expressing two recombinant single-chain variable fragment (scFv) antibodies, respectively scFvB9 and scFvH10. ScFvB9 was raised against glycoprotein G1 of Tomato spotted wilt virus (TSWV), and scFvH10 was raised against human tumor-associated antigen tenascin-C. Both antibodies were targeted to the secretory pathway using the N-terminal signal peptide from Phaseolus vulgaris polygalacturonase-inhibiting protein (PGIP), and scFvH10 carried in addition a C-terminal KDEL tetrapeptide for retention in the endoplasmic reticulum (ER). Sterile hydroponic cultures were established, allowing us to investigate whether scFvB9 and scFvH10 were present in root exudates. Intercellular fluids extracted from different plant tissues were analyzed by western blotting revealing the presence of scFvB9. Successful secretion of scFvB9 in hydroponic medium was also demonstrated, whereas no scFvH10 could be detected in the leaf, stem or root apoplast, nor secreted into the hydroponic medium. Our results show that scFvH10 release or diffusion from the roots of transgenic plants was not occurring, suggesting that the KDEL signal might contribute to the environmental biosafety of crops producing PMPs.
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PMID:Investigating recombinant protein exudation from roots of transgenic tobacco. 1908 Oct 9

Fungi are important natural product sources that have enormous potential for the production of novel compounds for use in pharmacology, agricultural applications and industry. Compared with other natural sources such as plants, fungi are highly diverse but understudied. However, research on Cladosporium cladosporioides revealed the existence of bioactive products such as p-methylbenzoic acid, ergosterol peroxide (EP) and calphostin C as well as enzymes including pectin methylesterase (PME), polygalacturonase (PG) and chlorpyrifos hydrolase. p-Methylbenzoic acid has ability to synthesise 1,5-benzodiazepine and its derivatives, polyethylene terephthalate and eicosapentaenoic acid. EP has anticancer, antiangiogenic, antibacterial, anti-oxidative and immunosuppressive properties. Calphostin C inhibits protein kinase C (PKC) by inactivating both PKC-epsilon and PKC-alpha. In addition, calphostin C stimulates apoptosis in WEHI-231 cells and vascular smooth muscle cells. Based on the stimulation of endoplasmic reticulum stress in some types of cancer, calphostin C has also been evaluated as a potential photodynamic therapeutic agent. Methylesterase (PME) and PG have garnered attention because of their usage in the food processing industry and significant physiological function in plants. Chlorpyrifos, a human, animal and plant toxin, can be degraded and eliminated by chlorpyrifos hydrolase.
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PMID:Cladosporium cladosporioides from the perspectives of medical and biotechnological approaches. 2833 73

The presence of articulated laticifers in the Moraceae family was recently discovered, which means that the location of pectinase and cellulase activities must be of great importance for their growth. Thus, the present study aimed to determine the role of these enzymes in the laticifer growth in Ficus montana and Maclura tinctoria. Reproductive meristems were collected and fixed in Karnovsky. Pectinase and cellulase labeling was performed in part of the samples, while another part was processed for usual TEM analyses. Pectinase and cellulase activities were detected in the vacuole and close to the middle lamella in both species. The presence of cellulases in the laticifers supports their articulated origin. Therefore, the occurrence of pectinase and cellulase activity in the laticifers points out that these enzymes could act in the dissolution of the transverse walls and in the processes of intrusive growth (through the dissolution of the middle lamella) and cell elongation (through the partial disassembly of components of the wall making it more plastic). Both enzymes are synthesized in the endoplasmic reticulum and transported to the cell wall by exocytosis or stored in the vacuole. The species studied showed a diverse subcellular composition, which is probably related to the species and not to the laticifer type (they present the same type) and to the composition of the latex (they show similar latex composition). We conclude that the presence of pectinases and cellulases can be used as a diagnostic condition for the laticifer types (articulated vs. non-articulated).
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PMID:Cellulases and pectinases act together on the development of articulated laticifers in Ficus montana and Maclura tinctoria (Moraceae). 3092 84

To investigate the molecular basis of multiple-allele-inherited male sterility in Chinese cabbage (Brassica campestris L. ssp. pekinensis), we performed differential proteomic analysis using iTRAQ to identify differentially abundant proteins between fertile and sterile flower buds from the genetic male sterile line 'AB01'. We identified 5932 high-confidence proteins; 1494 were differentially abundant between the two samples, including 749 up- and 745 down-regulated proteins. The up- and down-regulated proteins that could be essential for anther development and male sterility in sterile buds were mainly involved in (1) carbohydrate and energy metabolism (pyruvate dehydrogenase, glycolysis/gluconeogenesis, TCA cycle, starch and sucrose metabolism), (2) pollen wall synthesis and regulation (pectinesterase, polygalacturonase, pectate lyase, beta-galactosidase, glycosyl hydrolase), (3) protein synthesis and degradation (proteasome subunits, ribosome proteins, ABC transporters, RNA transport, protein processing in endoplasmic reticulum), (4) flavonoid biosynthesis, and (5) plant hormone signal transduction. We identified 10 genes/proteins that were both up-regulated and 122 that were both down-regulated in a conjoint analysis. Multiple reaction monitoring and qRT-PCR validation showed that the iTRAQ results were accurate and reliable. These findings will provide valuable information on proteins involved in anther development, and will contribute to the understanding of the molecular mechanism(s) that underlie male sterility in Chinese cabbage. BIOLOGICAL SIGNIFICANCE: Chinese cabbage is an allogamous plant with bisexual flowers that displays significant heterosis. The application of male sterile lines is a very efficient way to produce hybrid seeds, which can generate stronger plants that develop more rapidly and produce higher yield. However, the molecular mechanism(s) underlying multiple-allele-inherited male sterility in Chinese cabbage is unknown. In this study, we used a quantitative proteomic approach (iTRAQ) to identify DAPs between fertile and sterile buds of the GMS line 'AB01'. Subsequently, we also performed conjoined analysis of the iTRAQ results and our previously reported transcriptomics results. The aim of this research was to obtain the key DAPs and to identify the significantly enriched pathways involved in anther development and male sterility. These results may provide new insights into the molecular mechanism(s) underlying multiple-allele-inherited male sterility in Chinese cabbage.
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PMID:iTRAQ-based proteomic analysis of fertile and sterile flower buds from a genetic male sterile line 'AB01' in Chinese cabbage (Brassica campestris L. ssp. pekinensis). 3114 48