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Characterisation of PSC1 as an acidic rich RS domain protein (ARRS) with a conserved mammalian family member.Kavanagh, Steven James January 2006 (has links)
Title page, table of contents and summary only. The complete thesis in print form is available from the University of Adelaide Library. / The Acidic Rich family of RS Domain proteins (ARRS) is defined by both the presence and arrangement of conserved domains within 2 family members. Conserved regions include an RS domain, zinc finger domain, RNA binding motif and a C terminal acidic rich region. Two conserved motifs within the RRM of ARRS proteins have been defined that are not found in the RRM of other RNA binding proteins. Peri implantation stem cell 1 (pscl), the founding member of this family, was originally identified as a developmental marker differentially expressed between the Inner Cell Mass and primitive ectoderm of the mammalian embryo. Psc1 RNA is differentially up-regulated in the post gastrulation embryo and the adult, with high mRNA levels in lung, brain and kidney, and low level expression in other tissues. Comparative analysis of Psc1 to RS domain proteins known to function in mRNA processing, such as SC35 and ASF/SF2, has shown it colocalises to characteristic nuclear speckles. However, in contrast to SR proteins, Pscl localises to additional regions in the nucleus, not containing SR proteins, and to punctate regions in the cytoplasm termed cytospeckles. Further, in the absence of transcription, Psc 1 localises to regions in the nucleus which exclude nuclear speckles. Finally, unlike SC35 and ASF/SF2, which move rapidly in and out of nuclear speckles, FRAP assays show Psc1 is tethered within the nucleus. Analysis of Psc1 domain contribution to subcellular localisation and mobility shows the RRM to be both necessary and sufficient for Psc 1 cytospeckle localisation and is responsible for the nuclear tethering of Pscl. The RS domain of Pscl acts as a nuclear localisation signal and contributes to nuclear speckle localisation. The C terminal of Psc I localises with microtubules and is proposed to mediate Psc 1 cytoskeletal association. The expression of the Drosophila acidic rich RS domain protein (NP609976) is developmentally regulated in the same manner as Pscl, it has a nuclear localisation profile identical to Psc 1 and also localises to speckles in the cytoplasm, all of which support a conserved evolutionary role for Pscl and the ARRS protein family in mRNA processing and trafficking both in the nucleus and in the cytoplasm. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1236604 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 2006
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Chemical and Biochemical Factors That Influence the Gelation of Soybean Protein and the Yield of TofuBlazek, Vladimir January 2008 (has links)
Doctor of Philosophy(PhD) / Soybeans contain around 40% of high quality protein and 20 % of oil. Soy protein has long been used as ingredients for its emulsification and texturizing properties in a variety of foods, soymilk and tofu being the most popular. Soymilk is essentially a water extract of soybeans and there are many variations on the basic soymilk processing steps. Tofu, or bean curd, is made by coagulating soy milk, and then pressing the resulting curds into blocks. This thesis was mainly devoted to thermal denaturation and coagulation of soy proteins and targeted several selected important factors as they relate to the functional properties. The effects of different chemical coagulants as well as proteases on yield and quality of tofu from soybeans were studied. Eight tested chemical coagulants were able to coagulate the soymilk and the results showed that the concentration of soymilk and type of coagulant had a great influence on the properties of the tofu gel. The results also confirmed that the use of a suitable concentration of the quick-acting coagulants is more critical than that of the slow-acting coagulants in tofu making. In general, the extent of soymilk gelation is not determined by a single characteristic but rather results from a combination of factors. The gelation ability of various most common commercially available proteases to coagulate non-defatted soymilk was surveyed and the thermal stabilities of selected protease systems were compared. The difference in the temperature where the enzyme shows its highest activity seemed to be the most significant indicator when choosing a suitable enzyme for a certain industrial application. The three most effective and versatile soymilk coagulants were identified. The presence of small amounts of ficin in the system increased the protein recovery when calcium chloride was used as a coagulant. The most commonly used techniques of analysis of degree of hydrolysis (TNBS, OPA and pH-stat) of soy protein were compared. It was concluded that the pH-stat technique was useful for evaluating the progress of an enzyme-catalyzed protein hydrolysis process on an industrial scale while the OPA method seemed to be the most suitable method to be used for determining DH during the proteolysis of soymilk in laboratory conditions. The roles of soybean proteins, protein fractions and subunits to differences in gelling properties of different soybean varieties were examined. The variability and the interrelationship between soybean seed traits were established and the seed characteristics related to soymilk yield and tofu quality were identified. The results suggested that it is useful to predict the quality of tofu from a combination of characteristics of the soybean seed. It was concluded that large differences exist in soybean seed characteristics and their contributions towards the properties of the final product and implications were made towards the relative importance of individual soybean seed traits to the functional and textural properties of soy products. The SDS gel capillary electrophoresis was applied to characterize soybean storage proteins. The lab-on-a-chip technology was compared with capillary electrophoresis and these two methods were used to quantify the relative amount of 7S and 11S fractions in various soybean cultivars. It was concluded that both lab-on-a-chip instrument and a traditional CGE were adequate for analysis of soy-based products. Both systems were able to reliably quantify the relative amount of protein fractions in samples and thus demonstrate their different genetic origin. The great advantage of the lab-on-a-chip technology is its time-efficiency while the traditional CGE is a preferred instrument for method development. The usefulness of the chemometrical analysis of electrophoretic profiles as a method for objective evaluation, data reduction and interpretation was shown. The possibility of improvement of the protein extraction from soybeans in order to provide a basis for the optimization of soymilk production was studied. The enzyme-assisted extraction using the hydrolytic enzyme treatment to disrupt the soybean cell wall components was expected to improve the protein extraction yield. The results confirmed that the right selection of operational variables led to an increased yield of soymilk as well as its protein concentration. It was also shown that the addition of selected enzyme preparations into the soymilk process design resulted in an increased extraction yield of proteins from seeds into soymilk. The protein quality did not deteriorate during the enzyme-assisted extraction process and a small amount of microbial transglutaminase added together with a coagulant produced tofu with a significantly increased yield while maintaing satisfactory textural properties.
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Characterisation of the benzimidazole-binding site on the cytoskeletal protein tubulinLouisa M. MacDonald January 2003 (has links)
The binding kinetics of several benzimidazole compounds were determined with recombinant tubulin monomers and heterodimers from benzimidazole-sensitive and -insensitive organisms. This study utilised the naturally occurring high efficacy of the benzimdazoles for the parasitic protozoa Giardia duodenalis and Encephalitozoon intestinalis. The benzimidazoles are not active against the protozoan Cryptosporidium parvum or mammalian hosts, including humans. The affinity of several benzimidazole derivatives for monomeric and heterodimeric â-tubulin was clearly demonstrated, thus supporting previous studies of drug-resistant nematode and fungal populations. A homology model of protozoan áâ-tubulin, produced using the three-dimensionalstructure of mammalian áâ-tubulin, identified a strongly hydrophobic domain only on the â-tubulin protein of sensitive protozoa. This domain is proposed to be the benzimidazole-binding domain and the amino acid residues within it include three key residues which are substituted between benzimidazole-sensitive and insensitive organisms. These residues are Ile-189, Val-199, and Phe-200 that all have non-polar, hydrophobic side groups and are proposed to bind with the R5 side chain of several benzimidazole derivatives. In addition to this, the benzimidazole derivatives were able to bind irreversibly with assembling microtubules from sensitive parasites. The incorporation of benzimidazole-bound áâ-heterodimers into assembling microtubules was shown to arrest polymerisation in vitro although the addition of benzimidazole compounds to assembled microtubules did not result in depolymerisation. Taken together, these results suggest that the mechanism of action of these compounds is through disruption of the dynamic equilibrium that balances the cycle of microtubule polymerisation and disintegration within these protozoa. Further, this effect is brought about by preferential binding of the benzimidazoles to a hydrophobic region on the â- tubulin protein.
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Diffusion-collision model calculations of protein folding /Beck, Christopher A. January 2001 (has links)
Thesis (Ph.D.)--Tufts University, 2001. / Adviser: David L. Weaver. Submitted to the Dept. of Physics. Includes bibliographical references (leaves 148-149). Access restricted to members of the Tufts University community. Also available via the World Wide Web;
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Protein folding computational studies /Vasilkoski, Zlatko. January 2003 (has links)
Thesis (Ph.D.)--Tufts University, 2003. / Adviser: David L. Weaver. Submitted to the Dept. of Physics. Includes bibliographical references. Access restricted to members of the Tufts University community. Also available via the World Wide Web;
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Endotheliale Wirkmechanismen von rekombinantem aktiviertem Protein C am Beispiel von Fractalkine, Transforming Growth Factor-beta 2 und Cyclooxygenase-2 /Schulze Nahrup, Adriane. January 2007 (has links)
Universiẗat, Diss., 2007--Giessen.
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Physicochemical and structural effects of the obligatory activator calcium on the fast-twitch skeletal muscle isoform of phosphorylase kinasePriddy, Timothy Shane, Carlson, Gerald M. January 2006 (has links)
Thesis (Ph. D.)--School of Biological Sciences. University of Missouri--Kansas City, 2006. / "A dissertation in molecular biology and biochemistry and cell biology and biophysics." Advisor: Gerald M. Carlson. Typescript. Vita. Title from "catalog record" of the print edition Description based on contents viewed Nov. 9, 2007. Includes bibliographical references (leaves 103-113). Online version of the print edition.
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Die essentielle Bedeutung des" Classical Transient Receptor Potential 6" (TRPC6)-Ionenkanals für die akute vaskuläre Hypoxiereaktion der Lunge : Untersuchungen an isolierten pulmonalarteriellen glatten Muskelzellen /Fuchs, Beate. January 2007 (has links)
Universiẗat, Diss., 2007--Giessen.
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Development of a coarse-grained protein model and molecular dynamics studies of amyloid-[beta] peptide aggregation /Han, Wei. January 2007 (has links)
Thesis (Ph.D.)--Hong Kong University of Science and Technology, 2007. / Includes bibliographical references. Also available in electronic version.
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Membrane-assisted isoform immunoassay : separation and determination of protein isoforms /Lönnberg, Maria, January 2002 (has links)
Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser.
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