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21.
Genetic tailoring of N-linked oligosaccharides: the role of glucose residues in glycoprotein processing of Saccharomyces cerevisiae in vivo 总被引:1,自引:0,他引:1
In higher eukaryotes a quality control system monitoring the folding state
of glycoproteins is located in the ER and is composed of the proteins
calnexin, calreticulin, glucosidase II, and UDP-glucose: glycoprotein
glucosyltransferase. It is believed that the innermost glucose residue of
the N- linked oligosaccharide of a glycoprotein serves as a tag in this
control system and therefore performs an important function in the protein
folding pathway. To address this function, we constructed Saccharomyces
cerevisiae strains which contain nonglucosylated (G0), monoglucosylated
(G1), or diglucosylated (G2) glycoproteins in the ER and used these strains
to study the role of glucose residues in the ER processing of
glycoproteins. These alterations of the oligosaccharide structure did not
result in a growth phenotype, but the induction of the unfolded protein
response upon treatment with DTT was much higher in G0 and G2 strains as
compared to wild-type and G1 strains. Our results provide in vivo evidence
that the G1 oligosaccharide is an active oligosaccharide structure in the
ER glycoprotein processing pathway of S.cerevisiae. Furthermore, by
analyzing N- linked oligosaccharides of the constructed strains we can
directly show that no general glycoprotein glucosyltransferase exists in S.
cerevisiae.
相似文献
22.
Signe Sveegaard Jonas Teilmann Jakob Tougaard Rune Dietz Kim N. Mouritsen Geneviève Desportes Ursula Siebert 《Marine Mammal Science》2011,27(1):230-246
The population status of harbor porpoises has been of concern for several years, and the establishment of Marine Protected Areas (MPAs) has been suggested as a method to protect the harbor porpoise (Phocoena phocoena, Linneaus 1758) and other small cetaceans. In order to designate MPAs, high‐density areas for the species must be identified. Spatial distribution of small cetaceans is usually assessed from ship or aerial surveys. As a potentially more accurate alternative, this study examined the movements and area preferences of 64 harbor porpoises, satellite tagged between 1997 and 2007, in order to determine the distribution in the North Sea, the western Baltic, and the waters in between. Results show that harbor porpoises are not evenly distributed, but congregate in nine high‐density areas within the study area. Several of these areas are subject to significant seasonal variation. The study found no differences in the home range size of males and females, but immature harbor porpoises have larger home ranges than mature porpoises. The use of satellite telemetry for identifying areas of high harbor porpoise density can be of key importance when designating MPAs. 相似文献
23.
Ronny Martinez Felix Jakob Ran Tu Petra Siegert Karl‐Heinz Maurer Ulrich Schwaneberg 《Biotechnology and bioengineering》2013,110(3):711-720
Bacillus gibsonii Alkaline Protease (BgAP) is a recently reported subtilisin protease exhibiting activity and stability properties suitable for applications in laundry and dish washing detergents. However, BgAP suffers from a significant decrease of activity at low temperatures. In order to increase BgAP activity at 15°C, a directed evolution campaign based on the SeSaM random mutagenesis method was performed. An optimized microtiter plate expression system in B. subtilis was established and classical proteolytic detection methods were adapted for high throughput screening. In parallel, the libraries were screened for increased residual proteolytic activity after incubation at 58°C. Three iterative rounds of directed BgAP evolution yielded a set of BgAP variants with increased specific activity (Kcat) at 15°C and increased thermal resistance. Recombination of both sets of amino acid substitutions resulted finally in variant MF1 with a 1.5‐fold increased specific activity (15°C) and over 100 times prolonged half‐life at 60°C (224 min compared to 2 min of the WT BgAP). None of the introduced amino acid substitutions were close to the active site of BgAP. Activity‐altering amino acid substitutions were from non‐charged to non‐charged or from sterically demanding to less demanding. Thermal stability improvements were achieved by substitutions to negatively charged amino acids in loop areas of the BgAP surface which probably fostered ionic and hydrogen bonds interactions. Biotechnol. Bioeng. 2013; 110: 711–720. © 2012 Wiley Periodicals, Inc. 相似文献
24.
Hammes F Broger T Weilenmann HU Vital M Helbing J Bosshart U Huber P Odermatt RP Sonnleitner B 《Cytometry. Part A》2012,81(6):508-516
Accurate and sensitive online detection tools would benefit both fundamental research and practical applications in aquatic microbiology. Here, we describe the development and testing of an online flow cytometer (FCM), with a specific use foreseen in the field of drinking water microbiology. The system incorporated fully automated sampling and fluorescent labeling of bacterial nucleic acids with analysis at 5-min intervals for periods in excess of 24 h. The laboratory scale testing showed sensitive detection (< 5% error) of bacteria over a broad concentration range (1 × 10(3) -1 × 10(6) cells mL(-1) ) and particularly the ability to track both gradual changes and dramatic events in water samples. The system was tested with bacterial pure cultures as well as indigenous microbial communities from natural water samples. Moreover, we demonstrated the possibility of using either a single fluorescent dye (e.g., SYBR Green I) or a combination of two dyes (SYBR Green I and Propidium Iodide), thus broadening the application possibilities of the system. The online FCM approach described herein has considerable potential for routine and continuous monitoring of drinking water, optimization of specific drinking water processes such as biofiltration or disinfection, as well as aquatic microbiology research in general. 相似文献
25.
26.
Multiple structural alignment and clustering of RNA sequences 总被引:3,自引:0,他引:3
27.
Brianne E. Docter Scott Horowitz Michael J. Gray Ursula Jakob James C.A. Bardwell 《Nucleic acids research》2016,44(10):4835-4845
Organisms use molecular chaperones to combat the unfolding and aggregation of proteins. While protein chaperones have been widely studied, here we demonstrate that DNA and RNA exhibit potent chaperone activity in vitro. Nucleic acids suppress the aggregation of classic chaperone substrates up to 300-fold more effectively than the protein chaperone GroEL. Additionally, RNA cooperates with the DnaK chaperone system to refold purified luciferase. Our findings reveal a possible new role for nucleic acids within the cell: that nucleic acids directly participate in maintaining proteostasis by preventing protein aggregation. 相似文献
28.
29.
Jakob Parzefall Jacques P. Durand Bernard Richard 《Ethology : formerly Zeitschrift fur Tierpsychologie》1980,53(2):133-138
The chemical communication of the epigean Necturus maculosus and the cave-living Proteus anguinus are compared in laboratory conditions. In both species a specks-specific substance transferred by water could be shown, another was deposited on the substrate. 相似文献
30.