... upon treatment with the anticancer drugs doxorubicin and lovastatin [132] Among the three cancers, sarcoma cells possess the most Hsp25, the rodent equivalent of human Hsp27, andthe protein builds ... proteins and disease Therapeutic implications of sHSPs and other molecular chaperones Temperature induced synthesis of sHSPs protects against ischemia ⁄ reperfusion damage to the heart, brain, and ... and drugs The therapeutic application of sHSPs is further suggested by study of other molecular chaperones, with disruption of HSPs that protect deregulated intracellular signaling proteins and...
... Expression and purification of recombinant MTB HSP 16.3 in E coli Figure is the SDS/PAGE and Western immunoblot analyses ofthe expression and purification of MTB HSP 16.3 Induction of protein ... beginning at the time of heat shock, plated and CFU counted The proportions of viable cells expressing the pET 16b-MTB HSP 16.3 vector andthe two control vectors were plotted for 0, and h following ... measured with the addition of different concentrations of MTB HSP 16.3 and in the presence or absence of ATP and ATP analogs (A) Aggregation of CS is plotted in arbitrary units against time in the presence...
... peak at 7.65 mL on the elution profile Increase ofthe temperature of incubation was accompanied by the simultaneous increase ofthe peak eluted at 7.65 mL and decrease ofthe peak eluted at 8.65 ... polymerization of intact actin and aggregation of heated actin Interaction of HSP25 with intact actin Fig Effect of heating on the kinetics of polymerization (A) and saltinduced increase ofthe light ... (Fig 6A) Indeed the critical concentration of actin was equal to 0.25 ± 0.07; 0.21 ± 0.05, and 0.15 ± 0.06 lM in the absence of HSP25, and in the presence ofthe wild type HSP25 and its 3D mutant,...
... higher than in the absence ofthe sHsp This may reflect the formation of very large aggregates of MDH that also include the sHsp, as evidenced by the loss of sHsp from the SEC profile under these conditions ... composed of fewer than 12 subunits Size ofthe recombinant sHsp oligomers Although nondenaturing PAGE and SEC indicated the class I and II sHsps have different oligomeric structures, neither of these ... than the TaHsp17.8C-II particles, which have an estimated diameter of only nm Therefore, the relative sizes ofthe two oligomers are consistent with their behavior on nondenaturing PAGE and SEC Their...
... in a significant increase ofthe void volume fraction and a decrease of dimeric CS andthe second HspH peak (Fig 7A, broken line), indicative ofthe formation of large and stable chaperone–substrate ... indicated by the dotted lines assemblies A transfer of CS and HspH into the void volume, at the expense ofthe other fractions, was confirmed by SDS-PAGE (Fig 7C) Neither HspH nor CS shifted into the void ... the secondary structure The traces recorded are typical of b-strand proteins and show a single minimum elipticity in the range of 217 nm, similar to that observed for other sHsps [14,18,32] The...
... that the speckle staining is indeed due to the presence of phosphorylated aB-crystallin, we performed Western blotting with the anti(aB-crystallin) and anti-S45p IgGs on the isolated nuclei of these ... with the RIKEN mAb anti-(aB-crystallin) (a) or the monoclonal antibody to SC35 (d) and costained with anti-Sm (b and e) The yellow pseudocolour shows the extent of colocalization between the two ... as a percentage ofthe total aB-crystallin Values are based on four independent experiments and error bars represent the standard error ofthe mean (SEM) Asterisks indicate the aB-crystallin mutants...
... shown), and is a predominant residue in the flexible regions of both human and murine sHsps The majority of residues present in the flexible regions ofthe extensions (71% and 74% for human and murine, ... fluorescence depends strongly on the local environment ofthe amino acid and is a sensitive probe of conformation in the vicinity of tryptophan residues 5888 The binding of ANS and other hydrophobic probes ... [48] The hydrophilicity, lack of structure and associated flexibility are essential for the solubilizing role ofthe extension in the sHsp and complexation ofthe sHsp with target proteins The...
... between strands b2 and b6 of neighboring monomers The p26 modifications examined in this study are not located in either of these strands and they are not considered further As the basic p26 oligomer ... result ofthe residue’s location at the edge ofthe dimer–dimer interface The maximum oligomer size obtained with p26 R114A was 500 kDa, closer to the mass ofthe WT oligomer than the other variants ... tetramers, the next level of structure Modeling indicates that tetramer formation depends upon contact of b-strand 10 from the C-terminal extensions of monomers A and D with b-strands and in the a-crystallin...
... compare the apparent rates of trypsinolysis ofthe wild-type HSP22 and its mutants, we plotted ln(At ⁄ Ao) (where Ao and At are the intensities ofthe band of intact protein at the beginning of trypsinolysis ... respect to the location and length of specific b-strands For example, in the model of aB-crystallin, the b7 strand is only four residues long [16] whereas, in the model of HSP27, the same strand is ... However, the overall structures of aB-crystallin and HSP27 predicted by these two models are very similar, andthe positions ofthe b-strands correlate well with the corresponding positions of the...
... knockout of Hsf1 and Hsf4, the abnormal levels of FGF-7 returned to normal, and proliferation and differentiation ofthe epithelial cells were stabilized These findings indicate that HSF1 and HSF4 ... development ofthe lens and maintenance ofthe olfactory epithelium [102] The first evidence for a developmental function of HSF4 was provided by population genetic studies where mutations ofthe Hsf4 ... many cells and under different circumstances and that they are capable of interacting with each other The physical and functional interactions may therefore provide another layer of control for...
... distribution ofthe size ofthe inclusions was noticed Moreover, the percentage of cells presenting inclusions as well as the distribution ofthe size ofthe inclusions were dependent on when the analysis ... downstream ofthe initiator ATG in exon (Ex1) ofthe 67 exon-containing htt gene [5] Pathogenesis in HD correlates with the cleavage of mutated htt andthe release of an N-terminal fragment bearing the ... Note the presence ofthe granules in the cytoplasm ofthe cells Bar, 20 lm (D) The percentage of EGFP-containing cells displaying granules is shown The average percentages, including standard...
... p26-1Bam-s and p26-192Xhoas (Table 1) amplified only the cDNA in p26-full, and amplification of constructs with the primers employed for production of their respective inserts yielded PCR products ofthe ... transferred to nitrocellulose, and reacted with antibody to p26 using the ECL procedure The top of each gradient is to the left ofthe figure, and numbers across the top indicate successive samples ... gels, transferred to nitrocellulose, and reacted with antibody to the His6 epitope tag The top of each gradient is to the left ofthe figure, and numbers across the top indicate successive samples...
... (B) Rearrangement ofthe complexes formed by the 3D mutant of Hsp27 andthe wild-type Hsp20 A mathematical summation ofthe elution profiles ofthe isolated 3D mutant of Hsp27 and isolated wild-type ... independently of pH, a-crystallin was not precipitated and remained in the supernatant The changes of pH in the range of 6.0–7.5 weakly affect both amplitude andthe position of maximum on the far ... than that ofthe intact wild-type Hsp20 (Fig 1B) By analyzing the distribution ofthe charge residues in the C-terminal region of Hsp20, and by calculating the theoretical pI values of differently...
... of F-actin thermal denaturation All the methods used here show that the size of these complexes depends on the Hsp27-3D ⁄ actin ratio in the initial mixture of Hsp27-3D and F-actin (Figs 3, and ... together, all these results support a viewpoint that the formation of soluble complexes with non-native proteins is a conserved feature ofthe sHSP family of chaperones, andthe morphology of these ... means that the size ofthe first peak cannot be directly used for correct determination ofthe quantity of complexes formed by denatured actin and Hsp273D In contrast, the size ofthe second peak...
... intensities, with the peak of G and TS at 348 nm and R at 360 nm The fluorescence intensity of R was the lowest and it was red-shifted in comparison to other samples The extent of tertiary structure ... ANS and fluorescence was measured with an excitation wavelength of 388 nm and a band pass of nm, and an emission wavelength of 473 nm with a band pass of nm Measurements were carried out at either ... extension and another in the C-terminus The deletion of these motifs, termed G (multiple glycine), R (multiple arginine), and TS (multiple threonine ⁄ serine), was confirmed by sequencing andthe modified...
... as one ofthe three main consensus regions ofthe a-Hsp family [5] There is little experimental evidence to circumstantiate the role ofthe C-terminal extension, and even less concerning the conserved ... activity and oligomerization [44] In a-crystallin, on the other hand, numerous point mutations and even truncation ofthe first half ofthe N-terminal region did not affect complex formation and functionality ... isoleucine residues Two further constructs, HspH(D5C) and HspH(D15C), contained the entire I-X-I motif To assess the role ofthe conserved motif in the context ofthe full-length protein, three...
... only a handful of confirmed associations relative to the diversity of human neoplasms and pathogens Nevertheless, the worldwide population burden of infectious organisms makes understanding their ... receptors and increase the expression of IκBα, preventing the nuclear translocation ofthe pro-inflammatory molecule NF-κB [69,70] Heat-shock proteins and tumorigenesis The histologic evidence of chronic ... up-regulated in response to the development of high grade intraepithelial neoplasia and cervical cancer [92] These examples begin to unveil the complex relationship between HSPs andcancer formation Microbes,...
... measuring AC heating of Co- and Fe-ferrofluids with different viscosities Figure (b), and (c) show the dependence of surrounding viscosity of Co-ferrofluid, and Fe-ferrofluid on the AC magnetically-induced ... of Thesis Chapter presents the background, motivations, and objectives of this thesis Chapter reviews the previous works relevant to this project and discusses the theoretical background on the ... magnetic field of 178 Oe ~ 255 Oe and frequency of 1.2 MHz Since then, there have been a number of theoretical and experimental research activities to improve the magnetic hyperthermia [45] The systematic...
... investigate the role ofthe HSPs in antigen presentation and signaling Furthermore, we examine the immunogenicity and regulatory properties of HSPs using an in vivo mouse model In view ofthe issue of ... Srivastava and colleagues have made an important contribution to the discovery ofthe role of HSPs in cancer immunity, particularly in the processing and cross-presentation of antigen [Srivastava and ... However, due to the low levels of IL-2 secreted, the data is inconclusive The role of B pseudomallei Hsp70 and Hsp60 in the presentation ofthe short Ova peptide SIINFEKL To investigate whether Hsp70,...
... repressing factor ofthe heat shock response by regulating the expression of HSP-70 Both protein-protein interaction and catalytic activity ofthe PARP protein play a key role in modulation ofthe heat ... assays BZ, RA conceived the study RA, RLD participated in the design ofthe study All authors read and approved the final manuscript Competing interests The authors declare that they have no competing ... PARP-1 and poly (ADP-ribosylation) of HSF-1 reduce the availability of HSF-1 to initiate transcription The increase in HSF-1 content, albeit inactive with DIQ pretreatment further reinforces the...