plasmids used in this project

... Genotype of yeast strains used in this project 43 Plasmids used in this project 44 Compositions of SDS-PAGE 57 11 LIST OF ABBREVIATIONS aa amino acid Amp ampicillin CHO Chinese Hamster Ovary CPY ... Saccharomyces cerevisiae strains and plasmids used in this project are listed in Table and Table respectively 42 Table 1: Genotype of yeast strains used in this project Strain Genotype Source RH448 ... subset of the Vps class E proteins initiate ubiquitin dependent vacuolar protein sorting by selectively binding ubiquitinated cargo and directing sorting of these cargoes into MVB vesicles 1.5 VPS...

Ngày tải lên: 03/10/2015, 20:57

... achieved using existing LSA-PLUS mounting, and does not require expensive "19 inch" accommodation such as racks and cabinets When patching is required, it can be carried out reliably using latched ... patented point method of termination which outperforms the competition in ease of installation, quality, and durability of the connection • • • Insulation clamping ribs hold the wire firmly in position ... space used Meanwhile the routers, switches and hubs, can be located in a 19 inch rack in the air conditioned computer room, which is the ideal environment for the active equipment The looming of...

Ngày tải lên: 26/10/2013, 19:15

... crucial role in the regeneration of various tissues following injury [27] HGF is a mesenchymal cell-derived heparin-binding glycoprotein that is secreted as an inactive single-chain precursor ... normally remains inactive, probably associated with the extracellular matrix in the tissues producing it In response to tissue injury, such as hepatic and renal injury, the inactive single-chain HGF ... five human carcinoma cell lines: a lung carcinoma cell line A549, a colon carcinoma cell line LoVo, stomach carcinoma cell lines MKN45 and MKN74, and HepG2 Protease TMPRSS13 is inhibited by HAI-1...

Ngày tải lên: 15/02/2014, 01:20

... examined calpain 7–IST1 interactions by in vitro and in vivo binding experiments, using purified recombinant proteins and 4414 cultured mammalian cells expressing epitope-tagged proteins We also investigated ... recombinant human calpastatin domain (CSD1, calpain inhibitor protein), 0.5 lm ovocystatin (cysteine protease inhibitor protein), 20 lm MG-132 (proteasome inhibitor), 20 lm antipain (cysteine protease ... the MIT domain of VPS4 forms three-helix bundles ESCRT-III proteins commonly contain conserved amino acid sequences for MIT binding, termed MIT-interacting motif (MIMs), in their C-termini Two types...

Ngày tải lên: 18/02/2014, 04:20

... the non-TK1 like kinases deoxycytidine kinase [15], deoxyguanosine kinase and Drosophila melanogaster multi-substrate kinase [16] This indicates a different evolutionary origin of the two classes ... assayed for thymidine kinase activity under standard assay conditions with 100 lm dThd Thymidine kinase assay Thymidine kinase activity was assayed by measuring initial velocities using the DE-81 ... maintaining a proper balance between the dNTPs In addition to the ribonucleotide reductase-controlled pathway, the dTTP level is controlled by thymidine kinases and TMP nucleotidases, forming...

Ngày tải lên: 18/02/2014, 12:20

... OR2AG1 in vitro PDZ domain interactions have been well characterized and modes of binding have been grouped into three main classes of PDZ binding motifs, occurring at the C-terminus of the interacting ... present in the PDZ binding motifs of other membrane proteins [17] We also found interaction of PDZ domains + with ORs showing no classical PDZ binding motifs, indicating that the understanding of ... are outlined (white asterisk) (D) PDZ domains + both interact with OR2AG1_GST in vitro Interaction assay using in vitro translated PDZ domains + 2, PDZ domains + 4, PDZ domain and PDZ domain with...

Ngày tải lên: 18/02/2014, 13:20

... family kinases cell cycle arrest, leading to an increase in chromosomal aberrations and cell death, indicating that phosphorylation of this site is important for integrity of the G1 checkpoint [13] ... sample contained 2.5 lg of total protein Values are the average of four samples with error bars indicating the SEM induced by treatment of the cells with the protein tyrosine phosphatase inhibitor ... Src family kinase inhibitor PP2 [20] In comparison, we also assayed the inhibition of purified recombinant kinases C-SRC, C-YES and p49WEE1 by PP2 using the 96-well plate format kinase assay p49WEE1...

Ngày tải lên: 18/02/2014, 18:20

... (Toxin Technology, Sarasota, FL, USA) was made up in normal saline solution The SEC1 used in this study contained £ 25 pgÆmL)1 endotoxin because none of the SEC1 solutions induced gelation in ... continuously for 12 after injecting mL of 10 lm lucigenin (Sigma Chemical Co.) in HBSS into the stainless cell of the system The area under the curve was calculated to obtain total chemiluminescence ... also inhibited the SEC1-induced febrile response in rabbits The detection of NF-jB protein in the nucleus became apparent after 30 and increased dramatically up to 24 h (Fig 2A) The DNA-binding...

Ngày tải lên: 19/02/2014, 00:20

... MAR-binding proteins In this view, the direct association between DNA-PK and S ⁄ MAR-DNA or -binding proteins is a considerable point An experimental observation seems to be the case that in an in ... distribution on individual DNA is determined Interestingly, rodent cells that are defective in Ku80 or DNA-PKcs are incomplete in the repair of chromatin loops containing multiple DSBs, which were induced ... proteins, including nucleolin, Ku80, nucleolinD, HMGB1 and HMGB2, were determined with an Applied Biosystems 492 model sequencer according to the instructions For proteins that might be modified in...

Ngày tải lên: 19/02/2014, 06:20

... allow TFIIIB binding in an active state (i.e capable of interacting with TFIIIC to Gly stabilize binding) in the case of tRNA1 -1 and in an inactive state (incapable of efficient interaction with ... TFIIIB-containing fraction (0.3 m KCl eluate from heparin–Sepharose) was predicted to bind to the probe As a positive control TBP binding to this element was also included in the binding assays ... via interactions with TFIIIC The stable binding (heparinresistant complex formation) also required the presence 5196 of TFIIIC (Fig 3A) Independent binding of TFIIIB was again confirmed using...

Ngày tải lên: 20/02/2014, 02:21

... matrix/adhesion protein, possibly necessary for development of a normal fetus and in regulating immune responses within the intestine Induction of fgl2 in macrophages in response to viruses results in the ... (containing Oct-1), )77 to )71 (containing Sp1), )68 to )57 (containing Ets/Stat3) all significantly reduced fgl2 promoter activity The linker-scanning mutant )77 to )71, which contained the Sp1 binding ... same as the mutated sequences in Table used in the luciferase assay Ets core GGAA is italicized to indicate its overlapping with consensus Stat3 binding site underlined Name fgl2 fgl2 fgl2 fgl2...

Ngày tải lên: 20/02/2014, 11:20

... (MnSOD) The ratio of metals incorporated into SOD mutants examined showed a reduction in selectivity in the order: no glutamine > one glutamine[wt] > two glutamines > one glutamine (the double mutants) ... H31, is involved in FeSOD but not in MnSOD Only in MnSOD is the active site glutamine (Q146) also in proximity to the bound azide Azide binding could therefore be affected by electrostatic interactions ... achromatically as is the case for SOD in this system (Fig 2D) This aberrant staining has been observed before with high protein concentrations in the system used [24] Enzyme activity and metal...

Ngày tải lên: 21/02/2014, 01:21

... chromatin Fig Identification of chromatin binding domains in the N-terminal region of LBR and analysis of the binding mode Empty columns: four kinds of GST fusion proteins including N-terminal domains ... participate in the binding of vesicles However, surface remodeling of chromatin through initial interactions between NEP-B78 containing vesicles and chromatin may permit LBR-chromatin binding activity ... Chromatin binding assay (I): a method involving soluble proteins This method was used in the experiments for Fig A cytosol fraction of Xenopus eggs was boiled for 10 min, cooled in ice-water for min,...

Ngày tải lên: 22/02/2014, 04:20

... TNFa-induced U937 cell apoptosis by maintaining mitochondrial integrity and function [43], and ricin-induced apoptosis of U937 cells by maintaining an intracellular reducing condition by acting ... factor) binds to the Jun domain, and that the binding is dependent on the amount of glucocorticoid [13] In addition, this binding results in exhibition of anti -in ammatory effects through inactivation ... mRNAs containing Mn-SOD increase to counterbalance these phenomena The induction and increase of oxidative stress during pregnancy has been indicated by increasing lipid peroxidation in pregnant...

Ngày tải lên: 22/02/2014, 07:20

... Ab1–42, this toxicity was enhanced In the presence of homocysteine, Ab1–40 caused a 40% increase in toxicity over that with homocysteine alone, while Ab1–42 caused a further 50% increase in toxicity ... levels in the 0.1–0.25 mM range In these studies, homocysteine was found to elicit a dose-dependent increase in toxicity in VSMC In the presence of Ab peptides, this toxicity is exacerbated This ... levels are further increased in the presence of homocysteine DISCUSSION In CAA, Ab deposition occurs principally in association with the vascular basement membrane [8,12,47] Binding to the extracellular...

Ngày tải lên: 22/02/2014, 07:20

... *** *** Visfatin/cyclophilin mRNA Control GW1929 RSG Visfatin/cyclophilin mRNA Visfatin/cyclophilin mRNA Visfatin/cyclophilin mRNA A gene [23], was also induced to a similar extent in a dose-dependent ... visfatin could bind and activate insulin receptors [14] However, the insulinmimetic actions of visfatin are still debated [13] All these data suggest that visfatin might be a player linking several ... over-expressing visfatin displayed a 15–25% increase in total intracellular NAD+ levels [38] By using the pharmacological visfatin inhibitor, FK866, a significant decrease in the intracellular...

Ngày tải lên: 06/03/2014, 22:21

... mature enzyme in the direction opposite to that of substrate binding, thereby serving as a linker between the ‘hook’ domain and the N-terminus of the mature enzyme This N-terminal–linker–‘hook’ ... to this pH results in inactivation and unfolding of mature cathepsin B [22] Therefore, inactive unfolded cathepsin B was used as a negative control As expected, procathepsin B migrated as a single ... cysteine cathepsins, were shown to induce a conformational change in procathepsin B upon binding, resulting in propeptide removal from the active site cleft and conversion of the zymogen into...

Ngày tải lên: 07/03/2014, 03:20

... linked to, but distinct from, apoptosis [9,10] Beclin 1, a class III phosphatidylinositol 3-kinaseinteracting protein [11], plays a role in promoting autophagy [12] Beclin contains a Bcl-2-binding ... Bcl-2-binding domain which may serve as a point of cross-talk between the autophagic and apoptotic pathways Recently, a BH3 domain in the Bcl-2-binding domain of Beclin was shown to bind to Bcl-XL ... -XL-binding domain We next investigated the control of Beclin and its binding partner, Bcl-2, on autophagic activity Beclin was the first mammalian protein described to mediate autophagy [12] Beclin...

Ngày tải lên: 07/03/2014, 09:20

... constitutive, inducible and developmentally regulated genes (reviewed in [51]) GAF binds to GA-rich sequences and specifically interacts with the trinucleotide GGA via a single zinc-finger DNA binding domain ... novel class of regulatory proteins, and it contains a DNA binding domain conserved in two invertebrate developmental regulators, Erect Wing and P3A2 [17] Erect Wing is essential for the Drosophila ... [24,25] Interestingly, the expression of the genes encoding mtSSB and Pol c-b is transcriptionally regulated by the DNA replication-related-element binding factor (DREF) Indeed, in Drosophila this...

Ngày tải lên: 07/03/2014, 16:20

... unique N-terminal extension linked to the DNA-binding domain (DBD), hinge region and ligand-binding domain (LBD) of the AR The restricted expression pattern, the inhibitory effect on AR function ... Scintillation counting was performed in a 1500 Liquid Scintillation Analyzer (Packard) Acknowledgements We are indebted to Prof G Stock and Dr U.-F Habenicht for continuous interest in this project We ... underlined (B) DNA and deduced amino acid sequences of exon 1B The open reading frame is shown in bold capital letters The flanking intron regions are in italics (C) Localization of exon 1B in the...

Ngày tải lên: 07/03/2014, 16:20