abeta clearance in m alzheimer

... formation of huntingtin-containing inclusion bodies in Huntington’s disease encourages cell survival, whereas monomers and small inclusion bodies of huntingtin, a protein possessing abnormal ... invasiveness, reduces matrix metalloproteinases in vitro and eliminates production of avb3 integrin, a protein missing in normal melanocytes but often manufactured during the invasive phase [141] ... III intermediate filament proteins, desmin and vimentin, differing from previously described aggresomes because ubiquitin is absent and formation is microtubule-independent These HeLa cell inclusions...

Ngày tải lên: 19/02/2014, 18:20

... showing a protective effect of MTB HSP 16.3 against thermal killing in E coli Fig Sequence alignment of recombinant MTB HSP 16.3 and recombinant human aB-crystallin Amino-acid sequence alignment ... chymotrypsin taken at 5-min intervals over a 30-min period MTB HSP 16.3 was readily degraded by chymotrypsin and nearly all intact protein was degraded by 15 (A) In the presence of 3.5 mM ATP, MTB ... of recombinant MTB HSP 16.3 and recombinant human aB-crystallin Lane 1, molecular mass markers; lane 2, MTB HSP 16.3; lane 3, aB-crystallin In the Western immunoblot, the IT-4 antibody to MTB HSP...

Ngày tải lên: 23/03/2014, 21:21

... fact may indicate that the above-mentioned small domain with molecular mass 9–11 kDa may include subdomains and of actin After heating at 43 °C, actin turns into a state that is different from both ... actin polymerization In this case, polymerization was initiated by the addition of only KCl and MgCl2 (50 mM and mM, respectively) In this case increase in fluorescence was sigmoidal in time and ... actin was purified according to Pardee and Spudich [21] G-actin in buffer G (5 mM Tris/HCl, pH 8.2, 0.2 mM ATP, 0.1 mM CaCl2, 0.5 mM b-mercaptoethanol, mM NaN3) was stored on ice and used within...

Ngày tải lên: 20/02/2014, 23:20

... similarity in the N-terminal arm, suggesting substrate specificity is determined by the N-terminal arm The N-terminus of PsHsp18.1.C-I was also implicated in substrate interactions in bis-ANS binding ... 30 samples show a detectable complex peak that elutes earlier (7 min), which may represent some kind of intermediate in complex assembly As the vast majority of complexes elute in the column void ... described by Lee et al [15] using 0.6 lM MDH and purified sHsps from 0.18 to 3.0 lM (monomer) in 50 mM NA/Pi buffer, pH 7.5 Samples were incubated in mL quartz cuvettes in a thermostated water bath at...

Ngày tải lên: 07/03/2014, 15:20

... column (Amersham Pharmacia) and eluting at 0.4 mLÆmin)1 with 200 mM ammonium acetate, at °C Fractions corresponding to the major eluting peak were pooled and concentrated to 1.4 mgÆmL)1 using ... resuspended in binding buffer [500 mM KCl, 20 mM Tris/HCl, mM imidazole, 10% (v/v) glycerol, pH 7.9] containing mM phenylmethanesulfonyl fluoride and 10 lgÆmL)1 DNase I Cells were lysed in a French ... a-crystallin domain (named after the human lens protein, a-crystallin) of HspH The G114A protein was wild-type-like in terms of complex formation and heterooligomerization, but impaired in chaperone...

Ngày tải lên: 23/03/2014, 12:20

... phosphate buffered saline Equal numbers of about 106 cells were resuspended in 50 lL ice-cold lysis buffer [10 mM Tris pH 7.5, 100 mM KCl, mM dithiothreitol, mM EDTA, mM MgCl2, mM phenylmethanesulfonyl ... taken up in 100 lL buffer (10 mM Tris/HCl pH 7.8, 10 mM NaCl, mM dithiothreitol, mM MgCl2, mM phenylmethanesulfonyl fluoride, supplemented with a protease inhibitor cocktail from Roche) and incubated ... aspartic acids Mimicking phosphorylation of aB-crystallin reveals a distinct nuclear staining To determine the subcellular localization of aB-crystallin mutants we performed indirect immunofluorescence...

Ngày tải lên: 23/03/2014, 13:20

... oligomerization of sHsps [57] Truncation from the C-terminus of aA-crystallin to remove the IXI motif renders the protein unable to form oligomers [58], indicating that interactions involving ... determined by A280 values, in 50 mm phosphate buffer (pH 7.3) containing 0.02% (w ⁄ v) NaN3 Samples were heated from 25 to 100 °C at °CÆmin)1 in mL quartz cuvettes with a cm path length in a ... remains elusive, there is evidence that the binding of target proteins occurs in the groove between monomers, and involves a b-sheet region located at the beginning of the a-crystallin domain...

Ngày tải lên: 07/03/2014, 04:20

... Oligomer mass range represents the smallest to largest oligomers observed, and maximum monomer number refers to the number of subunits in oligomers of maximum mass Oligomer mass range (kDa) Maximum ... of monomers A and D with b-strands and in the a-crystallin domain of monomers C and B, respectively (Fig 8) A more prominent dimer–dimer interface occurs with b-strand of monomer B interacting ... a-crystallin domain modifications promote translocation remains uncertain To summarize, analysis of individual amino acid substitutions, coupled with molecular modeling of protein structure, indicate...

Ngày tải lên: 07/03/2014, 12:20

... 10 mm NaCl, 0.1 mm EDTA, 0.1 mm phenylmethanesulfonyl fluoride and 30 mm 2-mercaptoethanol, were mixed with an equal volume of mm DMS in 400 mm triethanolamine ⁄ HCl (pH 8.0) and incubated at 37 ... mutants were heated in buffer C2 (20 mm Tris-acetate pH 8.0, containing 150 mm NaCl, 0.1 mm EDTA, 15 mm 2-mercaptoethanol and 0.1 mm phenylmethanesulfonyl fluoride) at 70 °C for Point mutations of the ... (from 10 lg to 200 lg) of the wild-type HSP22 or its mutants in 150 lL of buffer C1 (20 mm Trisacetate pH 7.6, containing 150 mm NaCl, 0.1 mm EDTA, 15 mm 2-mercaptoethanol and 0.1 mm phenylmethanesulfonyl...

Ngày tải lên: 23/03/2014, 07:20

... signalling pathway originating from the membrane is unclear [67] Furthermore, an impact of re-organization of membrane microdomains has been demonstrated both in vitro and in vivo using the membrane ... Common features among the HSF family members Similarly to most transcription factors, the members of the HSF family are modular proteins composed of functional domains (see figure in [4]) The most ... regulatory mechanisms steering the different members of the mammalian HSF family The family provides an excellent example of how proteins that share common functional domains and bind similar DNA...

Ngày tải lên: 18/02/2014, 04:20

... from complete [23], because the proteasome cannot digest polyglutamine sequences and release them during degradation of polyglutamine-containing proteins [24] This may interfere with proteasome ... manufacturer’s instructions Immune complexes were detected by chemiluminescence using the ECLtmsystem (Amersham Biosciences) Autoradiographs were recorded on X-Omat LS films (Eastman Kodak) Quantitative ... with 3.7% formaldehyde, pH 7.4, in NaCl ⁄ Pi Permeabilization was for 3.5 in 0.2% Triton X-100 Examination of samples was performed in a LSM510 laser scanning confocal Zeiss Huntingtin inclusions...

Ngày tải lên: 19/02/2014, 06:20

... a-crystallin is in kinetic competition with aggregation to stabilize a monomeric molten-globule form of a-lactalbumin Biochem J 354, 79–87 Marini, I., Moschini, R., Corso, A.D & Mura, U (2000) Complete ... multimerization mediated by phosphorylation–sensitive intermolecular interactions at the amino terminus J Biol Chem 274, 9378–9385 63 Kumar, L.V.S & Rao, ChM (2000) Domain swapping in human aA ... chemiluminescence (ECL) procedure (Amersham Pharmacia Biotech) following manufacturer’s instructions The p26 bands were scanned with a BioÁRad Model GS-670 Imaging Densitometer and analyzed in MOLECULAR...

Ngày tải lên: 22/02/2014, 04:20

... the insulin B-chain (Sigma) [15] All experiments were performed in buffer E (50 mM phosphate, pH 7.5, 100 mM Na2SO4) Insulin (6.5 mgÆmL)1), dissolved in 2.5% acetic acid, was added to the incubation ... remaining in the supernatant ( 0.5 mgÆmL)1) had a negative maximum at 220 nm (Fig 5A) After dialysis at pH 6.8, wild-type Hsp20 ( 1.0 mgÆmL)1) was predominant in the supernatant and the maximum ... column (Amersham-Pharmacia) equilibrated with buffer B (20 mM Tris/acetate, pH 7.6, 10 mM NaCl, 0.1 mM EDTA, 0.1 mM phenylmethanesulfonyl fluoride, 14 mM b-mercaptoethanol) and developed by a linear...

Ngày tải lên: 07/03/2014, 15:20

... 30 column using the ACTA-FPLC system (Amersham Pharmacia, Biotech Europe GmbH, Helsinki, Finland) The column was equilibrated with 30 mm Hepes ⁄ KOH (pH 7.3) containing 100 mm NaCl and mm MgCl2 ... cm)1 Monomeric G-actin in G buffer (2 mm Tris ⁄ HCl, pH 8.0, 0.2 mm ATP, 0.2 mm CaCl2, 0.5 mm b-mercaptoethanol, mm NaN3) was polymerized into F-actin filaments by the addition of MgCl2 to a final ... Prior to experiments, F-actin was diluted to a final concentration (from 0.25 to 3.0 mgÆmL)1) with 30 mm Hepes (pH 7.3), containing 100 mm NaCl and mm MgCl2 Recombinant human Hsp27-3D was cloned,...

Ngày tải lên: 16/03/2014, 06:20

... oligomer mass maximum refers to the mass of the largest oligomer Maximum monomer number refers to monomer number in the largest oligomer Oligomer mass Monomer mass (kDa) Maximum monomer number ... Additionally, in contrast to the situation with bacteria, the maximum monomer number for oligomers produced by G and WT p26 in mammalian cells was the same Maximum monomer numbers for oligomers of ... sequence motifs in the p26 N-terminal extension and another in the C-terminus The deletion of these motifs, termed G (multiple glycine), R (multiple arginine), and TS (multiple threonine ⁄ serine),...

Ngày tải lên: 30/03/2014, 20:20

... a-crystallin domain including either the N-terminal region or the C-terminal extension were constructed in a similar manner Mutations leading to single and double amino-acid exchanges were introduced into ... concentration of 0.5 mM, and grown for a further 2–3 h After harvesting, cells were resuspended in binding buffer (500 mM KCl, 20 mM Tris/HCl, mM imidazole, pH 7.9) containing mM phenylmethanesulfonyl ... washing buffer (500 mM KCl, 20 mM Tris/HCl, pH 7.9) containing increasing imidazole concentrations (5–50 mM) The imidazole concentration was finally raised to 250 mM in order to elute bound proteins...

Ngày tải lên: 31/03/2014, 21:21

... chronic inflammation has been shown to contribute to tumorigenesis by causing DNA damage, promoting neoangiogenesis and compromising tumor immunosurveillance mechanisms Inflammation, tumor immunity ... simultaneously stimulating both the innate and adaptive immune system [53] Gp96-activated DCs release proinflammatory cytokines resulting in the induction of an inflammatory response by the innate component ... as aromatic amines and polycyclic aromatic hydrocarbons, generating carcinogenic metabolites [102] An increased number of inflammatory cells in the urinary bladder of schistosomal patients may...

Ngày tải lên: 10/08/2014, 22:20

... Property Measurement System (PPMS) 55 3.3.5 Magnetic Property Measurement System (MPMS) 56 3.3.6 Measurement of AC Magnetically-Induced Heat Temperature 56 3.3.7 Dynamic Light Scattering System (DLS) ... heating temperature TB blocking temperature TEM transmission of electron microscope Tmax maximum temperature TTT transpupillary thermotherapy VSM vibrating sample magnetometer XRD X-ray diffraction ... Hyperthermia Applications” 11th Joint MMM-Intermag Conference, Washington, DC, USA (2010, 1, 18 ~ 22) Asahi Tomitaka, Hiroki Kobayashi, Tsutomu Yamada, Minhong Jeun, Seongtae Bae, Yasushi Takemura, “Magnetization...

Ngày tải lên: 09/09/2015, 10:08

... the marrow was flushed with complete R10 medium (RPMI-1640 supplemented with 10 % FBS, mM L-glutamine, 50 M 3-mercaptoethanol, 100 U/mL penicillin and 100 µg/mL streptomycin) using a syringe ... calf serum (Gibco BRL, Grand Island, NY), mg/mL L-glutamine (Sigma), 100 U/mL penicillin and 100 µg/mL streptomycin (Sigma) and ng/mL of granulocyte macrophage-colony stimulating factors (GM-CSF, ... gift from Dr Ronald Germain, NIH) The cells were cultured in complete RPMI 1046 medium supplemented with 10 % heat-inactivated foetal calf serum, mg/mL L-glutamine and 100 U/mL penicillin and...

Ngày tải lên: 09/10/2015, 11:06