functional expression of heterologous potassium channels in yeast

... under the control of the GAL10 promoter The induction of the yeast, by galactose containing media, results in overexpression of GAL4, in turn inducing an increase of the expression of the OR gene ... function of increasing amount of total proteins in membrane fractions from I7 yeast strain induced at 15 °C, and from yeast expressing SSTR2 as a control activity By taking advantage of structural ... the inherent difficulties associated with the expression of ORs in heterologous cell systems [10] A major hindrance to functional expression of ORs has been the tendency of ORs to be retained in...

Ngày tải lên: 07/03/2014, 16:20

... ¯ow rate of mLámin)1 The following gradients were used: incubations containing arbutin or salicin as substrate; 100% C initial, increasing D to 20% (10 min), 50% (15 min), 100% (5 min), held ... Naringenin-7-GlcRha (naringin) Hesperetin-7-GlcRha (hesperidin) Phloretin-7-Glc (phlorizin) Oleuropein Cyanidin-3-Glc (kuromanin) Cyanidin-3,5-diGlc Flavanones Dihydrochalcones Secoiridoids Anthocyanidins Ó FEBS ... glycosides, 17% A/83% B initial (1 min), increasing B to 90% (10 min), held at 90% (4 min) The column was re-equilibrated (5 min) in the appropriate starting solvent conditions following gradient development...

Ngày tải lên: 08/03/2014, 16:20

... presence of CMP and AMP (from FAD) GMP was not present in any Qor extract, indicating that the host strains did not incorporate Mo-MGD, or free GMP, into the cofactor binding domain of the Qor protein ... molybdenum¨ containing hydroxylases of quinoline, isoquinoline, and quinaldine In Molybdenum and Tungsten Their Roles in Biological Processes (Sigel, A & Sigel, H., eds), pp 481–537, Vol 39 of Met Ions ... protein content, and determination of the apparent Km and kcat values for quinoline The activity of Qor was determined spectrophotometrically by measuring the quinoline-dependent reduction of the...

Ngày tải lên: 17/03/2014, 10:20

... ) Protein expression of CaSR in rat PASMCs and PAs Western blotting with monoclonal CaSR-specific antibody revealed signal of apparent molecular seize of 130 kD in the protein lysates of cultured ... mature form of CaSR Immunofluorescence staining showed that CaSR proteins were observed in vessel walls of PAs and were located in the cytoplasm and plasmalemma of the PASMCs, as shown in other ... The role of the calcium sensing receptor in regulating intra-cellular calcium handling in human epidermal keratinocytes J Invest Dermatol 2007, 127:1074-1083 Zhang WH, Xu CQ: Calcium sensing receptor...

Ngày tải lên: 10/08/2014, 05:21

... design, especially on sizing and positioning of cooling channels With the same instrument, optimal processing conditions in the cooling operation was found, minimising functions linked to process quality ... tooling incorporating conformal heating channels for transfer moulding of elastomer compounds, Plastic Rubber Compos 30 (8) (2001) 384–388 [5] M Ring, et al., An investigation of effectiveness of ... software for prototyping and simulation The study is on going and hopefully would culminate in the suggestion of the level of proficiency required using virtual models in deciding moulding specifications...

Ngày tải lên: 12/02/2014, 22:20

... with increased ability to complete the physical performance tests and in carrying out ADL (Table 6) Informants were present in 1167 (97%) of the interviews Comparison of the physical health rating ... Years of life expectancy free of problems with PADL 12.32 13.43 7.81 8.33 Years of LE free of problems with all ADL % of total LE free of problems with PADL 93.3 91.2 % of total LE free of problems ... for paying attention to programmes aimed at preventing and postponing disability and dependency Such emphasis is essential for improving the quality of life of the elderly, even in developing countries...

Ngày tải lên: 14/02/2014, 06:20

... the MCPIP gene The coding sequence lacking the PIN domain was obtained in two steps First, the fragment of 423 bp situated in front of the PIN domain was amplified by PCR using forward primer 5¢CGGGATCCCGGAGTCTGAGCTATGAGTG-3¢ ... E185, D226, D244) in PIN domains involved in binding Mg2+ ions are conserved in all three sequences Other conserved residues are R263, V139 and N144 Determination of the MCPIP domain architecture ... shown) Of the identified proteins, 73% are proteins involved in mRNA stability, but there are also proteins involved in other cellular processes, such as actin Further studies characterizing proteins...

Ngày tải lên: 18/02/2014, 14:20

... may in uence the expression of HO-1 The dominant expression of HO-2 protein, in comparison with the low expression of HO-1 protein, in H146 small cell lung cancer cells is of particular interest ... the expression of HO-2 mRNA and protein and induced the expression of HO-1 mRNA and protein In contrast, the scrambled HO-2 siRNA did not affect the expression of HO-1 and HO-2 mRNAs and proteins ... determine the basal expression levels of HO-1 Effects of HO-1 or HO-2 short interfering RNA (siRNA) on the expression of HO-1 Fig Effects of SA and SnPP on cellular heme content in human cell lines...

Ngày tải lên: 19/02/2014, 05:20

... HO-2 expression in human cell lines We initially analyzed the effects of hypoxia on the expression of HO-1 and HO-2 in human cell lines of Reduced expression of heme oxygenase-2 A B Fig Effects of ... regions including a large intron of the HO-2 gene are required In summary, the present study has suggested that the reduced expression of HO-2 protein may contribute to the maintenance of intracellular ... expression in human cells under hypoxia In the present study, we have analyzed the effect of hypoxia on the expression levels of HO-1 and HO-2 in various types of human cell line, including erythroleukemia...

Ngày tải lên: 19/02/2014, 06:20

... distinctive structural features of algal [Fe]hydrogenases are also present in the C reinhardtii HydA2 amino-acid sequence [33–35], including the well-conserved C-terminal part (C-domain) that binds ... differences in the regulation of expression of the two genes Gene expression during anaerobic induction The coexpression of the HydA1 and HydA2 transcripts was studied by Northern blot analysis In Fig ... hypothesis will be investigated in the future Fig Heterologous and homologous expression of the HydA2 protein Western blots of IPTG-induced E coli total protein extracts overexpressing, HydA1 or HydA2...

Ngày tải lên: 20/02/2014, 11:20

... Identification of N-glycosylation sites in the gonadotropin-releasing hormone receptor: role in receptor expression but not ligand binding Mol Cell Endocrinol 107, 241–245 47 Nussenzveig, D.R., Heinflink, ... xTRHR3 and xTRHR2 This is particularly interesting considering the functional importance of the third intracellular loop and the C-terminal tail in receptor coupling and regulation The amphibian EL1, ... sites of xTRHR1 (cAMP/cGMP-dependent protein kinase) and xTRHR2 (tyrosine kinase) are indicated with dashed and solid lines Since TRH desensitized the receptor (data not shown), one dose of TRH...

Ngày tải lên: 21/02/2014, 03:20

... group of GHMP kinases in which proton transfer does not play a major role in the rate determining step of catalysis That five of the 10 disease-causing mutations resulted in insoluble protein in ... and yeast enzymes in its kinetic mechanism The most likely cause of this sort of mechanism is that ATP binding induces a conformational change in the enzyme, which creates a functional binding ... quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72, 248–254 26 Wang, W & Malcolm, B.A (1999) Two-stage PCR protocol allowing introduction of...

Ngày tải lên: 08/03/2014, 02:20

... region of 11 aminoacid residues, a single hydrophobic segment of 20 aminoacid residues and a large luminal catalytic domain (498 amino-acid residues) Comparison of the primary structure of this ... full length ORF of hST6Gal II cDNA or with the pFlag vector by means of LipofectAMINE-reagent plus Expression of NeuAca2– 6Gal was detected using digoxigenin-labelled S nigra agglutinin and an anti-digoxigenin-fluorescein ... binding sites found upstream exon B This finding would argue for a ubiquitous and low-level expression of exon B containing transcripts On the other hand, the presence of CREB and SREBP binding...

Ngày tải lên: 08/03/2014, 08:20

... granules in skin melanophores resulting in darkening of the skin [23] POMC is the major cargo protein in this cell type and during adaptation to a black background the amount of POMC mRNA is induced ... population of neuroendocrine cells, but also of PrPC transgene expression, providing an additional tool for studying PrPC function Fig Schematic of the biosynthesis and processing of GFP–PrPC in intermediate ... (University of Leuven, Belgium [56]), and against the protein-folding chaperones calnexin and binding protein BiP by K Geering (University of Lausanne, Switzerland [57]) Monoclonal anti-tubulin IgG...

Ngày tải lên: 16/03/2014, 14:20

... properties of NaV1.2 The increase in ionic currents induced by coexpression of the human b1B could result from an increase in the number of channels present in the membrane, No, an increase in the ... studying the mechanism of upregulation of functional sodium channels on the cell surface and increasing the rate of spontaneous firing in peripheral neurons after nerve injury It will be interesting ... indicated by solid thin lines (b1) and shaded thin lines (b1B), and the unidentified 3¢ untranslated region of b1B is indicated using the thin interrupted broken line The stop codon is indicated by an...

Ngày tải lên: 16/03/2014, 23:20

... Kamerling, J.P & Vliegenthart, J.F.G (1992) The Asn-linked carbohydrate chains of human Tamm-Horsfall glycoprotein of one male Novel sulfated and novel N-acetylgalactosamine-containing N-linked ... appears in Mol Microbiol 31, 397±398) 10 Mattila, P., Rabina, J., Hortling, S., Helin, J & Renkonen, R È È (2000) Functional expression of Escherichia coli enzymes synthesizing GDP-L-fucose from inherent ... for protein expression as well as enzyme activity Expression of the fusion proteins was analyzed by Western blot using antibodies (Invitrogen) against the c-Myc and FLAG epitopes Chemiluminescence...

Ngày tải lên: 17/03/2014, 11:20

... quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72, 248–254 Wang W & Malcolm BA (1999) Two-stage PCR protocol allowing introduction of multiple ... of disease Some of the mutants have only small changes in the kinetic parameters In the case of N34S and D103G, disease-causation may be explained by a combination of kinetic impairment and increased ... during purification Kinetic analysis of disease-causing mutations The steady-state kinetic parameters of each of the nine disease-causing mutant proteins were determined (Table 1) In general, little...

Ngày tải lên: 23/03/2014, 11:20

... shown in the inset in (B) and in inset in (D) These results are in agreement with recent findings of Thiboutot et al [36] of 22R-hydroxycholesterol conversion into 17-hydroxypregnenolone in cultured ... occurring in 7-DHC, could affect 7-DHP hydroxy derivatives generating vitamin D3-like compounds (Fig 8) Such putative conversion would explain the lack of increase in vitamin D3 concentrations in ... chloroform/methanol (1 : 1, v/v), and the associated radioactivity measured by scintillation counting Fig Expression of MLN64 protein (arrow) in human skin (left) and human and rodent skin cells...

Ngày tải lên: 23/03/2014, 13:20

... from the number of experiments in parentheses SS Fig Increase in the UCP3 signal as a function of increasing amounts of (A) human recombinant protein and (B) mouse recombinant protein Representative ... amounts of UCP3 protein and mRNA in the three muscles are comparable Effect of fasting on UCP3 protein expression We and others have shown that fasting induces upregulation of UCP3 mRNA expression in ... quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72, 248–254 Muzzin, P., Eisensmith, R.C., Copeland, K.C & Woo, S.L (1996) Correction of obesity...

Ngày tải lên: 24/03/2014, 04:21

... proteins In fact, it was reported that the linker region of P-gp interacts with other proteins such as RING nger protein (RNF2) [25] and both alpha-tubulin and beta-tubulin [26] Therefore, in order ... restores the inwardfacing state The linker region of P-gp can be assumed to connect the C-terminal helix of subunit A (shown in red in Fig 7) with the N-terminal elbow helix of subunit B (shown in purple ... in purple in Fig 7) in the MsbA dimer In the inward-facing state, there appears to be less interaction between the linker region and each subunit Functional role of the linker region in P-gp because...

Ngày tải lên: 29/03/2014, 23:20